title: Peripheral blood monocyte GSK-343 Rep3

city: Rouen

first: Sylvain

last: Fraineau

address: 22 Gambetta Boulevard

department: UMR Inserm U1096

country: France

institute: Université de Rouen Normandie

description:
Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits respectively for human (StemCell Technologies Cat#19669). Treated with 5uM of GSK-343.
GSK-343_Treated_Rep3

data_processing:
Reads were aligned to the human genome (hg38) with Hisat2 version 2.0.5 with default parameters and using the GRCh38 transcript assembly.
Reads were then filtered to keep only properly paired and algined reads using samtools version 1.8.
Transcript quantification was performed with the subread tool featurecounts version 1.6.4.
Differential expression analysis was perfomed with DESeq2 version 1.22.1 under R version 3.5.1. Genes with a corrected p-value < 0.05 were considered significant.
Genome_build: hg38
Supplementary_files_format_and_content: Text file contains normalized read counts for each gene quantified for each samples/replicates, the log2 fold change between samples and the adjusted p-values.

source_name: Peripheral blood monocytes

growth_protocol: ISOLATION: Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits for human (StemCell Technologies Cat#19669). Treated with vehicule DMSO. CULTURE: Monocytes were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, Cat# 41966-029) supplemented with 50 µM of 2mercaptoethanol (Gibco Cat#21985-023), 10% Fetal Bovine Serum (FBS) and Penicillin/Streptomycin (Sigma-Aldrich Cat#P4333) at a cellular density > 150 000 ¢/cm² for primary monocytes and > 200 000 ¢/ml for TIB-204.

extract_protocol:
Total RNA was extracted from at least 3 x 10^6 human monocytes treated or not with 5 µM GSK-343 for 72h using NucleoSpin RNA XS kit (Macherley-Nagel Cat# 740902) using RNeasy Mini Kit (Qiagen Cat#74104) including a genomic DNA digestion step with DNAse I (Qiagen Cat#79254).
Library construction was conducted following manufactuer's instructions for the TrueSeq mRNA kit.

treatment_protocol: Cells were treated for 72h with 5 µM GSK-343 or DMSO.

tag: disease state

value: Non Coronary Heart Disease

value: Peripheral Blood

value: Monocyte


title: HEK293, ICI, 1.5h, n2

status: Public on Apr 24 2023

type: SRA

city: Geneva

first: Miriam

last: Stoeber

state: GE

address: Rue Michel Servet 1

department: PHYM

country: Switzerland

institute: University of Geneva

library_source: transcriptomic

data_processing:
1.5h: Libraries generated for the 1.5 h treatment were sequenced using the Illumina NextSeq 500 or HiSeq 4000 devices with a single-end read length of 80-90 nucleotides yielding at least 3 millions mapped reads per sample. Sequences were mapped to the human genome GRC h38.12 using GENCODE v28 annotation with STAR v. 2.6.0c. Generated raw counts were normalized and genes with low expression were filtered out leaving 15,946 genes. Differential expression analyses and related statistics between control and treated conditions were conducted using the Limma-voom R package 3.38.2.
6h: libraries were sequenced using the Illumina HiSeq 4000 device with a single-end read length of 50 nucleotides yielding around 30 million mapped reads per sample. Fastq files were generated using Illumina bcl2fastq v. 2.20.0 and mapped to the human genome UCSC hg38 using STAR v. 2.7.0f. Aligned reads were quantified with the Python software htseq-count v. 0.9.1. Genes were filtered down to 14,467 after removal of low expressers and normalized using the R package EdgeR 3.28.1. Differential gene expression between control and DPDPE or ICI and ICI/SNC80 conditions was computed and statistics were performed using the General Linear Model, quasi-likelihood F-test (QL F-test) with FDR and Benjamini & Hochberg correction.
Assembly: GRC h38.12 (1.5h), UCSC hg38 (6h)
Supplementary files format and content: 1.5h: tab delimited text files including ensembl, hgnc, Log2FC, p-value, adj. p-value, description
Supplementary files format and content: 6h: tab delimited text file including hgnc, description, FC, p-value, adj. p-value, raw counts, normalized counts

molecule: total RNA

organism: Homo sapiens

source_name: HEK293 stably expressing DOR

growth_protocol: HEK293 cells were maintained in DMEM supplemented with 10%FBS and geneticin

extract_protocol:
Total RNA was extracted using the RNeasy Mini Kit (Qiagen), genomic DNA was removed using the RNase-free DNase kit (Qiagen) and samples analyzed using a bioanalyzer.
Multiplexed libraries for the 1.5 h samples were generated using the Lexogen QuantSeq 3' mRNA-Seq Library Prep Kit and for the 6 h samples using TruSeqHT Stranded Total RNA Library Prep protocol (Illumina).

treatment_protocol: HEK293 cells stably expressing DOR were treated with vehicle control or DPDPE 100 nM or SNC80 100 nM or ICI 174, 864 100 uM + SNC80 100 nM for 1.5h or 6h (ICI preincubated for 5 min)

tag: cell line

value: HEK293 stably expressing DOR

tag: cell type

value: Human embryonic kidney

tag: treatment

tag: time point


title: HEK293, ICI, 1.5h, n1


title: Peripheral blood monocyte GSK-343 Rep1

description:
Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits respectively for human (StemCell Technologies Cat#19669). Treated with 5uM of GSK-343.
GSK-343_Treated_Rep1

molecule: polyA RNA

tag: tissue


title: Heart 41 Day 2 H9

city: London

first: Richard

middle: James

last: Mitter

state: LONDON

address: 1 Midland Road

department: Bioinformatics & Biostatistics

country: United Kingdom

institute: The Francis Crick Institute

data_processing:
The nf-core rnaseq pipeline (release v3) was used to generate gene-level abundance estimates using RSEM-STAR (https://github.com/nf-core/rnaseq)
The pipeline was run with default options except for specifying the GRCh38 genome fasta, Ensembl release 95 GTF file and STAR aligner (--fasta Homo_sapiens.GRCh38.dna_sm.primary_assembly.fa --gtf Homo_sapiens.GRCh38.95.gtf --aligner star_rsem)
Assembly: GRCh38 with Ensembl release 95 transcript annotations
Supplementary files format and content: RSEM gene abundance estimates

source_name: pluripotent stem cell line

growth_protocol: Cells were routinely passaged as sigle cells for differentiation. When cultures reached 80% confluency cells were differentiated in monolayer following the left ventricle cardiomyocyte differentiation protocol.

extract_protocol:
RNA was isolated using the Qiagen Rneasy kit
KAPA mRNA (polyA) HyperPrep

treatment_protocol: Cells were collected at specific time points and therefore were subject to different treatments in keeping with the left ventricle differentiation protocol

value: pluripotent stem cell line

tag: day

tag: experiment number


title: Heart 32 Day 60 H9


title: sample 01006 genomic DNA from blasts at day 15 during course 1

type: genomic

city: Freiburg

first: Geoffroy

last: Andrieux

address: Breisacherstr 153

country: Germany

institute: University clinics Freiburg

hyb_protocol: Samples were hybridized according to manufacturer's protocol (Illumina).

scan_protocol: Samples were scanned according to manufacturer's protocol (Illumina).

data_processing:
preprocessRaw function from minfi (V1.36) R package
normalized data: Average Beta

label: Cy5 and Cy3

molecule: genomic DNA

source_name: blasts

label_protocol: Samples were labeled with Cy5 and Cy3 according to manufacturer's protocol (Illumina).

growth_protocol: Mononuclear cells were isolated out of AML patients' whole blood samples by Pancoll density centrifugation. AML blasts were enriched by automated magnetic sorting using anti-CD34 and -CD117 beads.

extract_protocol: Genomic DNA was isolated with the DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany).

treatment_protocol: AML patients were treated within the DECIDER trial (NCT00867672) with decitabine ((Decitabine/DAC treatment, 20 mg/m2 intravenous 1-hour infusion over 5 days, with add-on drugs valproic acid and/or all-trans retinoic acid [ATRA] added at day 6).

tag: gender

value: female

value: blasts

tag: course


title: Heart 32 Day 20 H9


title: Heart 41 Day 0 H9


title: Peripheral blood monocyte Control Rep3

description:
Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits respectively for human (StemCell Technologies Cat#19669). Treated with vehicule DMSO.
DMSO_Control_Rep3

value: DMSO-Vehicle-Control


title: Heart 32 Day 15 H9


title: sample 25005 genomic DNA from blasts at day 8 during course 1

value: male


title: sample 01001 genomic DNA from blasts at day 15 during course 1


title: sample 26004 genomic DNA from blasts at day 8 during course 1


title: sample 26002 genomic DNA from blasts at day 8 during course 1


title: Heart 32 Day 6 H9


title: sample 31001 genomic DNA from blasts at day 8 during course 1


title: HEK293, ISO, 1.5h, n1

value: ISO


title: Peripheral blood monocyte Control Rep2

description:
Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits respectively for human (StemCell Technologies Cat#19669). Treated with vehicule DMSO.
DMSO_Control_Rep2


title: HEK293, SNC80, 1.5h, n1

value: SNC80 100 nM


title: Peripheral blood monocyte Control Rep1

description:
Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits respectively for human (StemCell Technologies Cat#19669). Treated with vehicule DMSO.
DMSO_Control_Rep1


title: sample 14008 genomic DNA from blasts at day 8 during course 1


title: sample 16005 genomic DNA from blasts at day 8 during course 1


title: Heart 32 Day 4 H9


title: sample 14007 genomic DNA from blasts at day 8 during course 1


title: HEK293, DPDPE, 1.5h, n3

value: DPDPE 100 nM


title: HEK293, ICI, 1.5h, n3


title: sample 14001 genomic DNA from blasts at day 8 during course 1


title: Heart 32 Day 2 H9


title: Peripheral blood monocyte GSK-343 Rep2

description:
Peripheral blood monocytes were selected by negative selection using EasySep™ monocyte isolation kits respectively for human (StemCell Technologies Cat#19669). Treated with 5uM of GSK-343.
GSK-343_Treated_Rep2


title: HEK293, DPDPE, 1.5h, n2


title: sample 04012 genomic DNA from blasts at day 8 during course 1


title: Heart 30 Day 60 H9


title: sample 11011 genomic DNA from blasts at day 8 during course 1


title: HEK293, SNC80, 1.5h, n3


title: HEK293, DPDPE, 1.5h, n1


title: HEK293, CTRL, 1.5h, n2


title: HEK293, SNC80, 1.5h, n2


title: Heart 30 Day 40 H9


title: HEK293, CTRL, 1.5h, n3


title: sample 03005 genomic DNA from blasts at day 8 during course 1


title: Heart 30 Day 20 H9


title: sample 01888 genomic DNA from blasts at day 8 during course 1


title: sample 01999 genomic DNA from blasts at day 8 during course 1


title: Bulk Transcriptome

city: Essen

first: Jürgen

middle: C

last: Becker

state: NRW

address: Universitätsstraße 1

department: Translational Skin Cancer Research

institute: Deutsches Krebsforschungszentrum

description:
transcriptome RNA count matrix
NextSeq 500/550
HTG Whole Transcriptome

data_processing:
The resulting FASTQ files were processed towards a gene expression count matrix using the HTG EdgeSeq Reveal Software version 4.0.1.
Assembly: GRCh38
Supplementary files format and content: barcodes.tsv.gz: raw barcode x feature matrix from cellranger output, features.tsv.gz: raw barcode x feature matrix from cellranger output, matrix.mtx.gz: raw barcode x feature matrix from cellranger output.
Transcriptome.csv: samples x genes matrix with unnormalized gene counts from EdgeSeq Reveal output; celltype_metadata.csv: Table with barcodes identified as cells and respective celltype annotation used through the scripts.
celltype_metadata.csv: Barcode and respective celltype annotation

source_name: oral squamous cell carcinoma (primary tumor tissue)

extract_protocol: The bulk transcriptome was analyzed using a quantitative nuclease protection assay from the HTG Transcriptome Panel (HTP) according to the manufacturers protocol (Cat. No. HTG-001-224, HTG Molecular Diagnostics, Tucson, AZ, USA). Briefly, the tumor areas were macro-dissected from 4 µm FFPE sections and subjected to Proteinase K and DNase digestion. Next, the quantitative nuclease protection assay was performed using the HTG EdgeSeq Processors before adapters and sample tags were added in a PCR amplification. The resulting libraries were sequenced using on an Illumina NextSeq 500/550 High Output Kit v2.5 (75 cycles) (Cat. No. 20024906, Illumina, Berlin, Germany).

value: oral squamous cell carcinoma


title: N2-B

city: Riverside

first: Adam

middle: David

last: Norris

state: CA

address: 3401 Watkins Drive

country: USA

institute: University of California, Riverside

data_processing:
STAR
HTSeq
Assembly: Caenorhabditis_elegans.WBcel235
Supplementary files format and content: HTSeq gene counts

organism: Caenorhabditis elegans

source_name: Whole animal

extract_protocol:
Trizol
NEBNext

value: Whole animal

tag: genotype


title: fust-1-B


title: fust-1-C


title: sample 01777 genomic DNA from blasts at day 8 during course 1


title: fust-1-ceh-14-A


title: sample 01039 genomic DNA from blasts at day 8 during course 1


title: dsODN_only-rep2

city: Chicago

first: Tong

last: Wu

state: Illinois

address: 929 E 57th St. GCIS E335

institute: University of Chicago

description: GUIDE-seq

library_source: genomic

data_processing:
GUIDE-seq was analysed using an open-source guideseq software:
https://github.com/aryeelab/guideseq
Assembly: hg19
Supplementary files format and content: read density that can be used for visualization on genome browsers.
Library strategy: GUIDE-seq

source_name: embryonic kidney

growth_protocol: HEK293T cells were purchased from (ATCC, CRL11268) were cultured in DMEM (Gibco 11995) supplemented with 10% (v/v) fetal bovine serum (Gibco), 1% penicillin and streptomycin (Gibco, 15140).

extract_protocol:
Genomic DNA was isolated using Purelink Genomic DNA Mini Kit (Thermo Fisher, K1820)
1 µg gDNA was applied to fragmentation, end-repair, A-tailing, adapter ligation, and dsODN-specific amplification, following the published GUIDE-seq protocol.

value: embryonic kidney

value: Kidney epithelial

value: Wild type

value: Nucleofection

tag: ascas12f version


title: dsODN_only-rep1

description: GUIDE-seq


title: fust-1-ceh-14-B


title: Heart 30 Day 10 H9


title: N2-A


title: Heart 30 Day 6 H9


title: enAsCas12f-HEXA-rep2

description: GUIDE-seq

tag: guide rna_target

value: HEXA


title: Heart 30 Day 2 H9


title: Heart 32 Day 0 H9


title: Heart 30 Day 4 H9


title: sample 01033 genomic DNA from blasts at day 8 during course 1


title: fust-1-ceh-14-C


title: enAsCas12f-HEXA-rep1

description: GUIDE-seq


title: sample 01031 genomic DNA from blasts at day 8 during course 1


title: tdp-1-ceh-14-A


title: sample 01032 genomic DNA from blasts at day 8 during course 1


title: AsCas12f-v4.1-HEXA-rep2

description: GUIDE-seq


title: Heart 30 Day 0 H9


title: tdp-1-ceh-14-B


title: sample 01027 genomic DNA from blasts at day 8 during course 1


title: Heart 28 Day 60 H9


title: AsCas12f-v4.1-HEXA-rep1

description: GUIDE-seq


title: Heart 30 Day 15 H9


title: Heart 28 Day 40 H9


title: HEK293, CTRL, 1.5h, n1

value: HEK293 stably expressing DOR


title: tdp-1-ceh-14-C


title: sample 01025 genomic DNA from blasts at day 8 during course 1


title: AsCas12f-WT-HEXA-rep2

description: GUIDE-seq

value: WT


title: N2-C


title: Heart 28 Day 20 H9

value: pluripotent stem cell line


title: sample 01022 genomic DNA from blasts at day 8 during course 1


title: AsCas12f-WT-HEXA-rep1

description: GUIDE-seq


title: sample 01016 genomic DNA from blasts at day 8 during course 1

value: female


title: ceh-14-A


title: Heart 28 Day 15 H9


title: ceh-14-B


title: Heart 28 Day 10 H9


title: enAsCas12f-TP53_2-rep2

description: GUIDE-seq


title: sample 01015 genomic DNA from blasts at day 8 during course 1

value: blasts


title: ceh-14-C


title: Heart 28 Day 4 H9


title: enAsCas12f-TP53_2-rep1

description: GUIDE-seq


title: AsCas12f-v4.1-TP53_2-rep2

description: GUIDE-seq


title: Heart 28 Day 2 H9


title: tdp-1-A


title: sample 01012 genomic DNA from blasts at day 8 during course 1


title: sample 01010 genomic DNA from blasts at day 8 during course 1


title: AsCas12f-v4.1-TP53_2-rep1

description: GUIDE-seq


title: tdp-1-B


title: AsCas12f-WT-TP53_2-rep2

description: GUIDE-seq


title: Heart 28 Day 0 H9


title: tdp-1-C


title: Patient 1, scRNAseq

data_processing:
Processing from FASTQ files with sequenced reads towards the unfiltered count matrix (barcodes x genes) was performed using Cellranger Software Suite version 3.1.0 and the human reference genome build GRCh38, downloaded from 10x Genomics in version 3.0.0.
Cells were identified from the barcodes by evaluation of four quality criteria: (1) number of unique molecular identifiers (UMIs), (2) number of genes, (3) percentage of mitochondrial gene expression and (4) number of expressed housekeeper genes, derived from Tirosh et al. without the mouse gene PRPS1L3. Evaluating these quality criteria and their relationship was inspired by the best practices of Theis et al.. We defined cells by having a minimum number of 500 UMIs and excluded cells with mitochondrial gene expression above 10%. The count matrix was processed using Seurat version 4.0.1 and R version 4.0.5. Cells were normalized using SCtransform algorithm and the 3000 most variable genes were selected for principal component analysis (PCA) using a single value decomposition approximation implemented in Seurat. Based on the variance explained by each PC we chose the first 20 PCs for UMAP visualization and clustering based on a K-nearest neighbor (KNN) graph as implemented in Seurat using resolution 0.8 .Cell types were identified using marker gene expression and differential gene expression of clusters that were merged based on their proximity in the UMAP. Malignant cells were additionally identified by inferred CNVs using inferCNV software suite version 1.6.0.
Genome_build: GRCh38 v3.0.0. (version by 10x Genomics)
Supplementary_files_format_and_content: barcodes.tsv: raw barcode x feature matrix from cellranger output, features.tsv: raw barcode x feature matrix from cellranger output, matrix.mtx: raw barcode x feature matrix from cellranger output, celltype_metadata.csv: Table with barcodes identified as cells and respective celltype annotation used through the scripts; Abbreviations: CAF, cancer-associated fibroblasts; Dend, Dendritic Cell; Macro, Macrophage. Cells labelled with “Missclassified“ were identified as wrong celltype after subsetting the respective cell type (see Methods)

source_name: left-sided neck

growth_protocol: immediate processing after surgery on same day; transport in Miltenyi tissue storage solution at 4°C

extract_protocol:
The native lymph node metastasis was immediately dissociated into single cell suspension using the gentleMACS Dissociator (catalog nr.: 130-093-235, Miltenyi Biotec, Bergisch Gladbach, Germany) with program "h_tumor_01", followed by 2x program "h_tumor_02" in 4.7 ml RPMI 1640 (catalog No. P04-16500 , PAN-Biotech) and an enzyme mix consisting of 200µl Enzyme H, 100 µl Enzyme R and 25µl Enzyme A (catalogue #130-095-929 Miltenyi Biotec). Cells were washed three times with 0.05% bovine serum albumin (BSA) phosphate-buffered saline (PBS) and filtered with a 100 µm cell strainer.
10,000 cells of the single cell suspension were then barcoded and processed with the 10x Genomics Chromium v2.0 platform (10x Genomics, Leiden, Netherlands) using the Chromium Single Cell 5' Library & Gel Bead Kits (1000006) and Library Construction Kit (1000020) of version 2. Library preparation was performed according to the manufacturers protocol and subsequently sequenced on an Illumina HiSeq 4000 (Illumina, Berlin, Germany) at the Genomics and Proteomics Core Facility of the german cancer research center (DKFZ, Heidelberg, Germany). Sequencing was performed in paired-end with 26 cycles on the forward read and 74 cycles on the reverse read. A total of around 353 million reads were sequenced.

value: lymph node metastasis from left neck

tag: tumor staging

value: pT1 pNx pMx R0 L0 V0 Pn1, ICD-O 8070/3

tag: Sex


title: sample 01007 genomic DNA from blasts at day 8 during course 1

value: male


title: AsCas12f-WT-TP53_2-rep1

description: GUIDE-seq


title: fust-1-A


title: M1 siSRSF12_R2

city: Porto

first: Joana

last: Wilton

address: Rua Alfredo Allen, 208

department: IBMC

country: Portugal

description: 3' RNA-Seq

data_processing:
Adapters were trimmed using Cutadapt v.1.13
Mapping to the hg38 genome was performed with topHat v.2 (ChrRNA-Seq) or with STAR (3'RNA-Seq).
Paired and mapped reads and bam file sorting were extracted using SAMtools v.
Bigwig (bw) foles were generated using the bamCoverage tool (Deeptools)
Genome_build: hg38
Supplementary_files_format_and_content: bigwig files: for all samples, we provide genome-coverage files

source_name: Primary human macrophages

growth_protocol: Macrophages were maintained in RPMI 1640 with 10% fetal bovine serum, with 1x penicillin/streptomycin and 50 ng/mL M-CSF; RKO andHCT15 were maintained in RPMI 1640 with 10% fetal bovine serum 1x penicillin/streptomycin

extract_protocol:
RNA was purified from primary human macrophages after the above timepoints, using Trizol and isopropanol precipitation
For ChrRNA-Seq, libraries were generated using NEBNext Ultra II Directional Prep Kit for Illumina (New England Biolabs), according to the NEB's protocol. For 3'RNA-Seq, libraries were generated using QuantSeq REV kit (Lexogen), according to Lexogen's instructions.

treatment_protocol: M0 macrophages were treated with RPMI 1640 with 2% fetal bovine serum for 24 hours; M1 macrophages were treated with LPS + IFN-gamma in RPMI 1640 with 2% fetal bovine serum for 24 hours and subsequently exposed to RKO or HCT15 for 24 hours. For transfection, primary human macrophages were polarised to M1 and then transfected with SiNTC (non-target control) or siSRSF12, in RPMI with 10% fetal bovine serum and 1x penicillin/streptomycin

tag: sample type

value: transfected with siSRSF12 siRNA, Dharmacon


title: M1 siSRSF12_R1

description: 3' RNA-Seq


title: sample 01001 genomic DNA from blasts at day 8 during course 1


title: M1 siNTC_R2

description: 3' RNA-Seq

value: transfected with siNTC siRNA, Dharmacon


title: sample 31001 genomic DNA from blasts at day 0 during course 1


title: sample 01004 genomic DNA from blasts at day 8 during course 1


title: M1 siNTC_R1

description: 3' RNA-Seq


title: sample 26004 genomic DNA from blasts at day 0 during course 1


title: sample 01013 genomic DNA from blasts at day 8 during course 1


title: M1+HCT15_R3

description: 3' RNA-Seq

value: co-cultured with cell line HCT15


title: M1+HCT15_R2

description: 3' RNA-Seq


title: sample 25005 genomic DNA from blasts at day 0 during course 1


title: sample 16005 genomic DNA from blasts at day 0 during course 1


title: M1+RKO_R3

description: 3' RNA-Seq

value: RKO


title: M1+HCT15_R1

description: 3' RNA-Seq


title: sample 14008 genomic DNA from blasts at day 0 during course 1


title: M1+RKO_R2

description: 3' RNA-Seq


title: sample 14007 genomic DNA from blasts at day 0 during course 1


title: sample 01006 genomic DNA from blasts at day 8 during course 1


title: M1+RKO_R1

description: 3' RNA-Seq


title: sample 11011 genomic DNA from blasts at day 0 during course 1


title: sample 14001 genomic DNA from blasts at day 0 during course 1


title: M1_R1

description: 3' RNA-Seq

value: pro-inflammatory


title: sample 26002 genomic DNA from blasts at day 0 during course 1


title: sample 03005 genomic DNA from blasts at day 0 during course 1


title: M1_R3

description: 3' RNA-Seq


title: sample 04012 genomic DNA from blasts at day 0 during course 1


title: sample 01999 genomic DNA from blasts at day 0 during course 1


title: M0_R2

description: 3' RNA-Seq

value: co-cultured with cell line RKO


title: M0_R3

description: 3' RNA-Seq


title: sample 01888 genomic DNA from blasts at day 0 during course 1


title: M0_R1

description: 3' RNA-Seq


title: sample 01777 genomic DNA from blasts at day 0 during course 1


title: sample 01039 genomic DNA from blasts at day 0 during course 1


title: OESO_0195

city: Brisbane

first: Katia

last: Nones

state: QLD

address: 300 Herston Road

country: Australia

institute: QIMR Berghofer Medical Institute

description: This submission

hyb_protocol: followed Illumina protocol

scan_protocol: Iscan following vendor instructions

data_processing: Raw idat files were imported using ChAMP (v.2.12.4).  Each data set, 450K and EPIC were imported and probes removed if the detected p-value >0.01 or or there were fewer than three beads in at least 5% of samples. Common probes to the two data sets (450K and EPIC arrays)  were normalized  using beta mixture quantile normalization (BMIQ) to account for probe type1 and type 2 biases, followed by a quantile normalisation (QN). Further filtering was performed to remove: 1) probes in non-CpG sites, 2) X or Y chromosome, 3) single-nucleotide polymorphism-related polymorphisms and 4) probes that map to multiple locations. A single value decomposition analysis determined a significant batch effect in the data sets. A batch correction was applied using removeBatchEffect from the limma package (v3.38.3) before any further analyses.

label: biotin

source_name: Tumour

label_protocol: followed Illumina protocol

extract_protocol: All prep DNA/RNA (Qiagen) following vendor protocol.

value: tumour


title: Heart 32 Day 40 H9


title: sample 01033 genomic DNA from blasts at day 0 during course 1


title: OESO_0272

description: This submission


title: sample 01032 genomic DNA from blasts at day 0 during course 1


title: Chrb M1+RKO_R2

description: ChrRNA-Seq


title: M1_R2

description: 3' RNA-Seq


title: OESO_0121

description: This submission


title: Chrb M1+RKO_R1

description: ChrRNA-Seq


title: OESO_0126

description: This submission


title: Chrb M1_R2

description: ChrRNA-Seq


title: Chrb M1_R3

description: ChrRNA-Seq


title: OESO_0120

description: This submission


title: sample 01027 genomic DNA from blasts at day 0 during course 1


title: sample 01025 genomic DNA from blasts at day 0 during course 1


title: RNA-seq T4 R2

city: Regensburg

first: Uwe

last: Schwartz

address: Universitätstraße 32

department: NGS Analysis Center

institute: University of Regensburg

data_processing:
STAR aligner was used to align the RNA-seq reads simultaneously to the human and the adenoviral genome. Read mapping was conducted using following parameters: --outSAMmultNmax 1, --outMultimapperOrder Random, --outFilterType BySJout, --outFilterIntronMotifs RemoveNoncanonical, --outSAMstrandField intronMotif, --outFilterMultimapNmax 20, --alignSJoverhangMin 8, --alignSJDBoverhangMin 1, --alignIntronMin 20, --alignIntronMax 1000000
To quantify the RNA abundance for each gene the number of uniquely mapped reads per gene was counted using the featureCounts function of the subread package
Genome_build: hg19 and AY339865.1 (adapted)
Supplementary_files_format_and_content: tab-delimited text files include raw count values for each sample

source_name: H1299 cells

extract_protocol:
Total cellular RNA was isolated from infected cells at 0, 0.5, 1, 2, 4 hpi using TRI Reagent (Molecular Research Center, Inc.). Two time series sets of total RNA were prepared from two independent infections.
Library preparation, comprising rRNA depletion, polyA enrichment, fragmentation to ~270 nucleotide length, reverse transcription to cDNA using random hexamer primers and adapter ligation, was accomplished using TruSeq RNA Sample Preparation Kit v2 (Illumina) according to the manufacturers protocol.

tag: hours post infection

value: 4 hpi


title: OESO_0119

description: This submission


title: sample 01016 genomic DNA from blasts at day 0 during course 1


title: RNA-seq T2 R2

value: 2 hpi


title: Chrb M1+RKO_R3

description: ChrRNA-Seq


title: sample 01015 genomic DNA from blasts at day 0 during course 1


title: RNA-seq T2 R1


title: OESO_0113

description: This submission


title: OESO_0075

description: This submission


title: OESO_0107

description: This submission


title: OESO_0106

description: This submission


title: OESO_0098

description: This submission


title: OESO_0118

description: This submission


title: sample 01013 genomic DNA from blasts at day 0 during course 1


title: OESO_0109

description: This submission


title: OESO_0125

description: This submission


title: OESO_0100

description: This submission


title: RNA-seq T1 R2

value: 1 hpi


title: OESO_0096

description: This submission


title: OESO_0074

description: This submission


title: OESO_0099

description: This submission


title: RNA-seq T1 R1


title: sample 01010 genomic DNA from blasts at day 0 during course 1


title: sample 01007 genomic DNA from blasts at day 0 during course 1


title: sample 01022 genomic DNA from blasts at day 0 during course 1


title: OESO_0070

description: This submission


title: infection 3h DMEM/F-12 cond biol rep 3

city: Uppsala

first: Staffan

middle: Gunnar

last: Svärd

address: Husargatan 3

department: Department of Cell and Molecular Biology

country: Sweden

institute: Uppsala University

description:
matrix column name:
259

data_processing:
Illumina NovaSeq 6000 , Demultiplexing and conversion to FASTQ performed using the bclfastq2 (2.20.0.422) software
FASTQ files were mapped to combined Giardia WB genome and human GRCh38 using STAR v2.7.10a. ReadsPerGene.out.tab output from STAR with specifying --quantMode GeneCounts were used as the raw counts input for downstream analysis
Gene differential expression analysis was carried out in R v4.0.1 using edgeR v3.32.1 workflow. Quasi-likelihood F test was used to determine significant differential gene expression.
Assembly: Giardia intestinalis WB C6,  GCA_000002435.2 UU_WB_2.1
Assembly: GRCh38, release 106, GCA_000001405.28
Supplementary files format and content: Tab-delimited text file containing count matrix of human raw counts from STAR mapping of all samples (counts_human.txt).
Supplementary files format and content: Tab-delimited text file containing count matrix of Giardia intestinalis raw counts from STAR mapping of all samples (counts_giardia.txt).

organism:
Homo sapiens
Giardia duodenalis assemblage A

source_name: trophozoites, jenjunal enterocytes

growth_protocol:
Giardia intestinalis trophozoites were grown in TYDK at 37°C until 70% confluency. Trophozoites were washed twice in warm PBS and after incubated in either DMEM/F-12 supplemented with 10% FBS or with DMEM/F-12 for 1h at 37°C. After this preconditioning, trophozoites were detached, spun down, washed once in 1ml cold DMEM/F-12, spun down again and after resuspended in DMEM/F-12.
Human enteroids were maintained as 3D domes embedded in Matrigel in human Intesticult organoid growth media in humidified atmosphere with 5% CO2 at 37°C. For infections enteroids were grown as monolayers on PET-transwells and differentiated to an enterocyte phenotype by deprivation of WNT signaling for 4 to 5 days. 3h before the infection apical media was changed to DMEM/F-12 while the basal compartment was refreshed with differentiation media.

extract_protocol:
Total RNA extractions were performed using Trizol according to the manufactures instructions. RNA was then treated with DNaseI and extracted again by phenol chloroform extraction.
Sequencing libraries were prepared from 500ng total RNA using the TruSeq stranded mRNA library preparation kit (cat# 20020595, Illumina Inc.) including polyA selection. Unique dual indexes (cat# 20022371, Illumina Inc.) were used. The library preparation was performed according to the manufacturers’ protocol.

treatment_protocol:
Giardia intestinalis trophozoites were added apically to enterocytes for 1h and 3h. Uninfected enterocyte monolayers were collected after 1h and 3h as controls. Prior to infection preconditioned Giardia intestinalis trophozoites were collected as inoculum samples.
All samples were collected and lysed in Trizol reagent.

value: trophozoites, jenjunal enterocytes

tag: time

value: DMEM/F-12 preconditioned, infected


title: OESO_0094

description: This submission


title: RNA-seq T0.5 R1

value: 0.5 hpi


title: OESO_P0065

description: This submission


title: sample 01006 genomic DNA from blasts at day 0 during course 1

value: female

value: blasts


title: RNA-seq T4 R1


title: sample 01031 genomic DNA from blasts at day 0 during course 1


title: RNA-seq T0.5 R2


title: RNA-seq T0 R2

value: 0 hpi


title: transfection reagent only 2

type: RNA

city: Kagoshima

first: Yuya

last: Tomioka

country: Japan

institute: Kagoshima University

hyb_protocol: 0.6 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a Agilent fragmentation buffer and was hybridized at 65°C for 17 hr using Gene Expression Hybridization Kit.

scan_protocol: Microarray slides were scanned at 3 μm resolution using an Agilent SureScan (G4900DA).

data_processing: The scanned images were analyzed with Feature Extraction Software (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded

source_name: mock 2

label_protocol: For cRNA amplification and labeling, Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies) were used following the manufacter’s instructions.

growth_protocol: Normal cell culture medium RPMI 1640

extract_protocol: Total RNA was extracted using Trizol following the manufacturer's instructions.

treatment_protocol: microRNA transfection using Lipofectamin RNAiMAX

value: lung adenocarcinoma

tag: cell line


title: infection 3h DMEM/F-12 cond biol rep 2

description:
matrix column name:
258

tag: cell type


title: sample 01012 genomic DNA from blasts at day 0 during course 1

value: male


title: sample 01004 genomic DNA from blasts at day 0 during course 1


title: miR-486-3p transfected cell

source_name: miR-486-3p transfected cell


title: RNA-seq T0 R1


title: transfection reagent only 1

source_name: mock 1


title: OESO_J0063

description: This submission


title: miR-486-5p transfected cell

source_name: miR-486-5p transfected cell


title: infection 3h DMEM/F-12 cond biol rep 1

description:
matrix column name:
257

tag: treatment


title: sample 01001 genomic DNA from blasts at day 0 during course 1


title: OESO_0087

description: This submission


title: low MNase-seq T4 R2

data_processing:
Paired-end reads were mapped to the reference using bowtie2-aligner with following paramters: --very-sensitive-local, --no-discordant
Reads were filtered to mapping quality > 30 and only reads mapped in proper pair were kept using samtools.
Fragments mapped to the Adenovirus reference with a maximum insert size of 139 bp were selected and used for pVII positioning analysis
pVII positions were called using the DANPOS2 toolkit. First, a pVII occupancy profile for each sample was generated and normalized to 20,000 reads using the dpos function with following parameters: -u 1e-5; -a 1; -z 5; -p 0.001, --extend 35 and --mifrsz 0. Next, the profiles were normalized by a genome wide quantile normalization to the first replicate at 0 hpi of each MNase condition (high/low) using the wiq script. Finally, pVII positions were obtained from the normalized pVII occupancy profiles using the dpos function with following parameters: -jd 50; -q 10; -z 5; -a 1.
Genome_build: hg19 and AY339865.1 (adapted)
Supplementary_files_format_and_content: biwig and bed files were generated using DANPOS2 toolkit.  bigwig files represent normalized pVII occupancy and bed files are called pVII positions

extract_protocol:
H1299 cells infected with Ad were digested at 0, 0.5, 1, 2 and 4 hpi with 100U/600U of MNase for 4/5 min at 37°, denoted as ‘low’/’high’ MNase. After fragmented DNA has been separated on 1.3% agarose gels, mono-nucleosomal fractions were isolated
Library preparation was accomplished using the NEBNext DNA library prep Master Mix Kit (Nex England Biolabs) according to the manufacturers protocol.

tag: mnase concentration


title: Chrb M1_R1

description: ChrRNA-Seq


title: infection 3h DMEM/F-12+FBS cond biol rep 3

description:
matrix column name:
256

value: DMEM/F-12+FBS preconditioned, infected


title: low MNase-seq T4 R1


title: infection 3h DMEM/F-12+FBS cond biol rep 2

description:
matrix column name:
255


title: infection 3h DMEM/F-12+FBS cond biol rep 1

description:
matrix column name:
254


title: low MNase-seq T1 R1


title: low MNase-seq T0.5 R2


title: infection 1h DMEM/F-12 cond biol rep 3

description:
matrix column name:
249


title: infection 1h DMEM/F-12 cond biol rep 2

description:
matrix column name:
248


title: OESO_P0054

description: This submission

tag: tissue


title: low MNase-seq T0.5 R1


title: infection 1h DMEM/F-12 cond biol rep 1

description:
matrix column name:
247


title: OESO_0062

description: This submission


title: infection 1h DMEM/F-12+FBS cond biol rep 3

description:
matrix column name:
246

value: DMEM/F-12+FBS preconditioned infected


title: high MNase-seq T4 R2


title: infection 1h DMEM/F-12+FBS cond biol rep 2

description:
matrix column name:
245


title: OESO_0048

description: This submission


title: OESO_0051

description: This submission


title: low MNase-seq T0 R1


title: infection 1h DMEM/F-12+FBS cond biol rep 1

description:
matrix column name:
244


title: OESO_0057

description: This submission


title: high MNase-seq T4 R1


title: high MNase-seq T2 R2


title: high MNase-seq T2 R1


title: OESO_P0052

description: This submission


title: high MNase-seq T1 R2


title: OESO_0038

description: This submission


title: pancreatic ductal adenocarcinoma cells, RalGAPβ-deficient-2, rep2

city: Tianjin

first: Mingxin

last: Cao

address: NO.22, Qixiangtai Road

country: China

institute: Tianjin medical university

description: Gene expression data from another clonal human pancreatic ductal adenocarcinoma cell line with RalGAPβ deficiency, obtained through RalGAPβ CRISPR-Cas9 treatment with indel mutation in RalGAPβ genomic region

hyb_protocol: Following fragmentation, cDNAs were hybridized for 16 h at 45℃ on Clariom S human array. Arrays were washed and stained using GeneChip Hybridazation, Wash, and Stain Kit.

scan_protocol: GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G3000 7G.

data_processing: The data were analyzed with RMA 2.0 using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 100.

source_name: MIA PaCa-2 cells, RalGAPβ-deficient clone-2

label_protocol: Biotinylated cDNAs were prepared according to the standard Affymetrix protocol from 500 ng total RNA (GeneChip WT PLUS Reagent Kit, Affymetrix).

growth_protocol: Cell lines were cultured in Dulbecco's modified Eagle medium containing 10% (v/v) fetal bovine serum, 100 U/ml penicilin, and 100 μg/ml strptomycin in a humidified atmosphere with 5% CO2 at 37℃.

extract_protocol: Total RNAs were extracted using Isospin Tissue & Cell RNA extraction kit (Nippon Genetics).

treatment_protocol: Two lines of RalGAPβ-deficient MIA PaCa-2 cells (KO1 and KO2) were established by the CRISPR-Cas9 system. A clonal cell lline with RalGAPβ  CRISPR-Cas9 treatment, but without indel mutation in the RalGAPβ genomic region was used as Control.

value: MIA PaCa-2

tag: cell line source

value: MIA PaCa-2 cells obtained from the American Type Culture Collection

value: RalGAPbeta KO2


title: OESO_0047

description: This submission

value: tumour


title: pancreatic ductal adenocarcinoma cells, RalGAPβ-deficient-2, rep1


title: pancreatic ductal adenocarcinoma cells, RalGAPβ-deficient-1, rep2

description: Gene expression data from a clonal human pancreatic ductal adenocarcinoma cell line with RalGAPβ deficiency, obtained through RalGAPβ CRISPR-Cas9 treatment with indel mutation in RalGAPβ genomic region

source_name: MIA PaCa-2 cells, RalGAPβ-deficient clone-1

value: RalGAPbeta KO1


title: high MNase-seq T1 R1


title: high MNase-seq T0.5 R1


title: high MNase-seq T0.5 R2


title: high MNase-seq T0 R2


title: low MNase-seq T1 R2


title: F5

city: Montreal

first: Gustavo

last: Turecki

state: Quebec

address: 6875 Boulevard LaSalle

department: Psychiatry

country: Canada

institute: McGill University

description: Batch: 6F, Chemistry:  v3

library_source: transcriptomic single cell

data_processing:
10X Genomics Cellranger v3.1.0 mkfastq
10X Genomics Cellranger v5.0.1 count
Cell filtering as described in publication
Clustering and cell type annotation: Unsupervised clustering of nuclei was performed with the Seurat algorithm, after using highly variable genes for PCA and correcting for batch effects with the Harmony R package. Clusters were annotated to cell types based on the enrichment of expression of known marker genes.
Differential expression analysis between cases and controls was performed separately for males and females within each broad cell type and cluster, using the summed counts of every gene per subject and with the muscat and edgeR R packages.
Assembly: refdata-gex-GRCh38-2020-A,   https://support.10xgenomics.com/single-cell-gene-expression/software/release-notes/build
Supplementary files format and content: count matrix for filtered genes and barcodes for all samples combined. This includes, in Matrix Market format, female samples whose raw data is provided here and male samples from the previous study. The matrix row names (genes) and column names (barcodes and cell-type annotations) are also given in CSV format.

source_name: Brain BA9 dlPFC

extract_protocol: Chromium Single Cell 3’ Reagent Kits v2 and v3.1

value: Brain BA9 dlPFC

value: Female

tag: group

value: Case


title: pancreatic ductal adenocarcinoma cells, no indel mutation, rep1

description: Gene expression data from a clonal human pancreatic ductal adenocarcinoma cell line obtained through RalGAPβ CRISPR-Cas9 treatment while without indel mutation in RalGAPβ genomic region

source_name: MIA PaCa-2 cells, no indel mutation

value: no indel mutation (control)


title: OESO_0027

description: This submission


title: OESO_0045

description: This submission


title: pancreatic ductal adenocarcinoma cells, no indel mutation, rep2


title: OESO_0046

description: This submission


title: high MNase-seq T0 R1


title: F19

description: Batch: 3F, Chemistry:  v2


title: Liver, T1D, exposure, rep3

status: Public on Apr 23 2023

city: Guangzhou

first: Wen

last: Chen

address: No. 74, 2nd Zhongshan Road

department: Department of Toxicology

institute: School of Public Health, Sun Yat-sen University

data_processing:
To process sequences, the reads were filtered to obtained using SOAnuke (https://github.com/ BGI-flexlab/SOAPnuke).
The clean reads were aligned to a reference genome using Bowtie2 (http://bow tie-bio.sourceforge.net/Bowtie2/index.shtml).
The FPKM method was used to calculate the unigene expression level by RSEM (http://deweylab.biostat.wisc.edu/ rsem/rsem-calculate-expression.html).
Assembly: GRCm38
Supplementary files format and content: xlsx files including FPKM values for each sample

organism: Mus musculus

source_name: Liver

extract_protocol:
The frozen tissues of lung and liver (approximately 5mg) were subjected to RNA isolation using TRIzol reagent (Invitrogen, Carlsbad, CA, USA).
Oligo dT magnetic beads were employed to trap mRNAs with poly A tails, and the mRNAs were fragmen- ted and reversely transcribed to double-stranded cDNA (dscDNA) by random primers. The cDNAs were ligated adaptors and subjected to amplification by PCR. The PCR products were then denatured, and single stranded PCR products were cyclized by splint oligos with DNA ligase to construct cDNA library.

value: Liver

value: WT

value: Type 1 diabetes, exposed to PM


title: OESO_0003

description: This submission


title: pancreatic ductal adenocarcinoma cells, no treatment, rep2

description: Gene expression data from a human pancreatic ductal adenocarcinoma cell line with no treatment

source_name: MIA PaCa-2 cells, no treatment


title: Liver, T1D, exposure, rep2


title: pancreatic ductal adenocarcinoma cells, no treatment, rep1


title: OESO_0035

description: This submission


title: Liver, T1D, exposure, rep1


title: OESO_0010

description: This submission


title: OESO_0011

description: This submission


title: Liver, ND, exposure, rep3

value: Normal diet feeding, exposed to PM


title: Liver, ND, exposure, rep2


title: Liver, ND, exposure, rep1


title: Liver, ND, control, rep3

value: Normal diet feeding, exposed to filtered air


title: Liver, ND, control, rep2


title: Liver, T1D, control, rep2

value: Type 1 diabetes, exposed to filtered air


title: Liver, ND, control, rep1


title: Liver, DIO, exposure, rep3

value: Diet-induced obesity, exposed to PM


title: Liver, DIO, exposure, rep2


title: Liver, DIO, exposure, rep1


title: Liver, T1D, control, rep1


title: Liver, DIO, control, rep2

value: Diet-induced obesity, exposed to filtered air


title: Lung, T1D, exposure, rep3

source_name: Lung

value: Lung


title: Liver, DIO, control, rep1


title: Lung, T1D, exposure, rep1


title: Lung, T1D, control, rep2


title: Liver, T1D, control, rep3


title: Lung, T1D, control, rep1


title: Lung, T1D, control, rep3


title: Lung, ND, control, rep2


title: Lung, ND, exposure, rep3


title: Lung, ND, control, rep1


title: Lung, T1D, exposure, rep2


title: Lung, ND, exposure, rep2


title: SUM159PT NNMT KOd A

city: Basel

first: Mohamed

last: Bentires-Alj

address: Hebelstrasse 20

department: DBM

institute: University Hospital Basel

description: SUM159PT NNMT KOd A

hyb_protocol: Bisulphite-treated samples were hybridized on Human Infinium Methylation EPIC BeadChip with standard Illumina protocol.

scan_protocol: Standard Illumina protocol.

data_processing: Illumina’s Genome Studio software V2.0

label_protocol: DNA was used for bisulphite conversion using the EZ-96DNA Methylation-Lightning™ MagPrep kit (D5046, Zymo Research).

growth_protocol: Cultures of SUM159PT NNMT WT, KOs and KOd cells for 3 days. Cells were grown in Ham's F12 (Sigma, cat#N6658) with 5% fetal calf serum (FCS) (Sigma, cat# F7524), 5 μg/mL human recombinant insulin (Sigma, cat# 91077C), 1 μg/mL hydrocortisone (Sigma, cat# H0888), 1× penicillin/streptomycin (Sigma, cat# P4333).

extract_protocol: Total genomic DNA from SUM159PT WT, KOs and KOd was isolated, including RNAse A treatment, according to the manufacturer protocol (Macherey Nagel, 740952.50).

tag: genetic background

value: KOd


title: Lung, ND, control, rep3


title: Lung, DIO, exposure, rep2


title: Lung, DIO, exposure, rep3


title: Lung, ND, exposure, rep1


title: Lung, DIO, control, rep3


title: GasN24_3

city: Chongqing

first: Min-Dian

last: Li

state: China

address: 30 Gaotanyan Main Street

department: Department of Cardiovascular Medicine

institute: Southwest Hospital, Army Medical University

description:
GasN24_3_1.fq.gz
GasN24_3_2.fq.gz

data_processing:
The sequencing data was filtered with SOAPnuke by (1) Removing reads containing sequencing adapter; (2) Removing reads whose low-quality base ratio (base quality less than or equal to 5) is more than 20%; (3) Removing reads whose unknown base ('N' base) ratio is more than 5%, afterwards clean reads were obtained and stored in FASTQ format.  SOAPnuke:Version v1.5.2  Parameters -l 15 -q 0.2 -n 0.05
The clean reads were mapped to the reference genome using HISAT2.   HISAT2: version: v2.0.4  Parameters：--phred64 --sensitive --no-discordant --no-mixed -I 1 -X 1000
Bowtie2 was applied to align the clean reads to the gene set. The expression level of gene was calculated by RSEM and represented as TPM.  Bowtie2: version: v2.2.5  Parameters: -q --phred64 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -I 1 -X 1000 --no-mixed --no-discordant  -p 1 -k 200  RSEM: version: v1.2.12
Genome_build: mm10/GRCm38.p6
Supplementary_files_format_and_content: tab-delimited text files include TPM values for each Sample
Supplementary_files_format_and_content: *csv: TPM and circadian parameters

source_name: adult female muscle

growth_protocol: Special pathogen-free (SPF) mice were purchased from Hunan SJT Laboratory Animal Co. and housed in a SPF barrier facility. C57BL/6J female mice at 7 weeks of age were grouped housed, and entrained to a 12h light:12h dark cycle with normal chow food and water ad libitum for 7 days before assigned to daytime-restricted feeding (DRF) and nighttime-restricted feeding (NRF) groups for additional 7 days.

extract_protocol:
Tissues were dissected, flash frozen liquid nitrogen, and RNA was harvested using Trizol reagent.
Oligo(dT)-attached magnetic beads were used to purified mRNA. Purified mRNA was fragmented into small pieces with fragment buffer at an appropriate temperature. Then First-strand cDNA was generated using random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis. afterwards, A-Tailing Mix and RNA Index Adapters were added by incubating to end repair. The cDNA fragments obtained from previous step were amplified by PCR, and products were purified by Ampure XP Beads, then dissolved in EB solution. The product was validated on the Agilent Technologies 2100 bioanalyzer for quality control. The double stranded PCR products from previous step were heated denatured and circularized by the splint oligo sequence to get the final library. The single strand circle DNA (ssCir DNA) was formatted as the final library. The final library was amplified with phi29 to make DNA nanoball (DNB) which had more than 300 copies of one molecular, DNBs were loaded into the patterned nanoarray and pair end 100 bases reads were generated on BGIseq500 platform (BGI-Shenzhen, China).

treatment_protocol: Animals were subjected to daytime-restricted feeding (DRF) or nighttime-restricted feeding (NRF) for 7 days

value: muscle

value: female

tag: strain

tag: age

value: 9 weeks

tag: dietary regimen

value: NRF

tag: zeitgeber time in hours


title: GasN24_4

description:
GasN24_4_1.fq.gz
GasN24_4_2.fq.gz


title: GasN24_2

description:
GasN24_2_1.fq.gz
GasN24_2_2.fq.gz


title: Lung, DIO, exposure, rep1

tag: genotype


title: Liver, DIO, control, rep3


title: Lung, DIO, control, rep1


title: SUM159PT NNMT KOs C

description: SUM159PT NNMT KOs C

value: KOs


title: GasN20_3

description:
GasN20_3_1.fq.gz
GasN20_3_2.fq.gz

tag: Sex


title: GasN20_4

description:
GasN20_4_1.fq.gz
GasN20_4_2.fq.gz


title: GasN20_2

description:
GasN20_2_1.fq.gz
GasN20_2_2.fq.gz


title: GasN20_1

description:
GasN20_1_1.fq.gz
GasN20_1_2.fq.gz


title: GasN16_3

description:
GasN16_3_1.fq.gz
GasN16_3_2.fq.gz


title: GasN16_4

description:
GasN16_4_1.fq.gz
GasN16_4_2.fq.gz


title: GasN16_2

description:
GasN16_2_1.fq.gz
GasN16_2_2.fq.gz


title: GasN16_1

description:
GasN16_1_1.fq.gz
GasN16_1_2.fq.gz


title: GasN12_2

description:
GasN12_2_1.fq.gz
GasN12_2_2.fq.gz


title: SUM159PT NNMT WT C

description: SUM159PT NNMT WT C


title: GasN12_4

description:
GasN12_4_1.fq.gz
GasN12_4_2.fq.gz


title: GasN08_4

description:
GasN08_4_1.fq.gz
GasN08_4_2.fq.gz


title: GasN12_1

description:
GasN12_1_1.fq.gz
GasN12_1_2.fq.gz


title: GasN08_3

description:
GasN08_3_1.fq.gz
GasN08_3_2.fq.gz


title: GasN08_2

description:
GasN08_2_1.fq.gz
GasN08_2_2.fq.gz


title: Lung, DIO, control, rep2


title: GasN04_4

description:
GasN04_4_1.fq.gz
GasN04_4_2.fq.gz


title: SUM159PT NNMT WT B

description: SUM159PT NNMT WT B


title: SUM159PT NNMT KOs B

description: SUM159PT NNMT KOs B


title: SUM159PT NNMT KOs A

description: SUM159PT NNMT KOs A


title: GasN04_3

description:
GasN04_3_1.fq.gz
GasN04_3_2.fq.gz


title: GasN04_2

description:
GasN04_2_1.fq.gz
GasN04_2_2.fq.gz


title: SUM159PT NNMT KOd B

description: SUM159PT NNMT KOd B


title: GasN12_3

description:
GasN12_3_1.fq.gz
GasN12_3_2.fq.gz


title: pancreatic ductal adenocarcinoma cells, RalGAPβ-deficient-1, rep1

value: MIA PaCa-2

value: MIA PaCa-2 cells obtained from the American Type Culture Collection

value: RalGAPbeta KO1


title: GasN04_1

description:
GasN04_1_1.fq.gz
GasN04_1_2.fq.gz


title: SUM159PT NNMT KOd C

description: SUM159PT NNMT KOd C


title: GasN00_4

description:
GasN00_4_1.fq.gz
GasN00_4_2.fq.gz


title: GasN00_2

description:
GasN00_2_1.fq.gz
GasN00_2_2.fq.gz


title: GasN08_1

description:
GasN08_1_1.fq.gz
GasN08_1_2.fq.gz


title: GasN00_3

description:
GasN00_3_1.fq.gz
GasN00_3_2.fq.gz


title: GasD24_1

description:
GasD24_1_1.fq.gz
GasD24_1_2.fq.gz

value: DRF


title: GasD24_2

description:
GasD24_2_1.fq.gz
GasD24_2_2.fq.gz


title: GasD24_4

description:
GasD24_4_1.fq.gz
GasD24_4_2.fq.gz


title: GasD24_3

description:
GasD24_3_1.fq.gz
GasD24_3_2.fq.gz


title: GasD20_4

description:
GasD20_4_1.fq.gz
GasD20_4_2.fq.gz


title: GasD20_2

description:
GasD20_2_1.fq.gz
GasD20_2_2.fq.gz


title: GasD20_1

description:
GasD20_1_1.fq.gz
GasD20_1_2.fq.gz


title: GasD20_3

description:
GasD20_3_1.fq.gz
GasD20_3_2.fq.gz


title: GasD16_4

description:
GasD16_4_1.fq.gz
GasD16_4_2.fq.gz


title: GasD16_2

description:
GasD16_2_1.fq.gz
GasD16_2_2.fq.gz


title: GasD16_1

description:
GasD16_1_1.fq.gz
GasD16_1_2.fq.gz


title: GasD12_2

description:
GasD12_2_1.fq.gz
GasD12_2_2.fq.gz


title: GasD16_3

description:
GasD16_3_1.fq.gz
GasD16_3_2.fq.gz


title: GasN00_1

description:
GasN00_1_1.fq.gz
GasN00_1_2.fq.gz


title: GasD08_2

description:
GasD08_2_1.fq.gz
GasD08_2_2.fq.gz


title: GasD12_4

description:
GasD12_4_1.fq.gz
GasD12_4_2.fq.gz


title: GasD08_4

description:
GasD08_4_1.fq.gz
GasD08_4_2.fq.gz


title: SUM159PT NNMT WT A

description: SUM159PT NNMT WT A

value: WT


title: GasD08_1

description:
GasD08_1_1.fq.gz
GasD08_1_2.fq.gz


title: GasD08_3

description:
GasD08_3_1.fq.gz
GasD08_3_2.fq.gz


title: GasD04_2

description:
GasD04_2_1.fq.gz
GasD04_2_2.fq.gz


title: GasD12_3

description:
GasD12_3_1.fq.gz
GasD12_3_2.fq.gz


title: GasD04_4

description:
GasD04_4_1.fq.gz
GasD04_4_2.fq.gz


title: GasD04_1

description:
GasD04_1_1.fq.gz
GasD04_1_2.fq.gz


title: GasD04_3

description:
GasD04_3_1.fq.gz
GasD04_3_2.fq.gz


title: GasD00_4

description:
GasD00_4_1.fq.gz
GasD00_4_2.fq.gz


title: GasD00_2

description:
GasD00_2_1.fq.gz
GasD00_2_2.fq.gz


title: GasD00_3

description:
GasD00_3_1.fq.gz
GasD00_3_2.fq.gz


title: GasD12_1

description:
GasD12_1_1.fq.gz
GasD12_1_2.fq.gz


title: GasD00_1

description:
GasD00_1_1.fq.gz
GasD00_1_2.fq.gz


title: WGBS_Day8plus_rep1

city: Maebashi

first: Tetsuro

last: Komatsu

address: 3-39-15 Showa-machi

institute: Gunma University

description:
Day8-DFO48h.cpg.cover.bedGraph.gz
Day8-DFO48h.cpg.methyl.bedGraph.gz

data_processing:
Reads were mapped on the mixed reference of mouse genome mm9 and lambda phage using BMap.
Alignments exported from BMap were summarized into a binary file in graph format using MPTC.
The methylation levels and coverage were exported from the graph file using MethExport.
Evaluation of CpG methylation levels within 1 kbp upstream from all TSSs was conducted using custom Perl scripts (available with test data at https://github.com/j-kohmaru-gunma/NGS_Program): First screening CpGs located within 1 kbp upstream from all TSSs and omitting CpGs having <5 or >1,000 coverage/read depth, then calculating changes in methylation levels from Days 0 to 8, and finally picking up genes having 3 or more CpGs that showed more than 50% reduction on methylation levels from Day 0.
DMR detection was performed using the software metilene (version 0.2-8, q value <0.05), and motif analysis for DMRs was carried out using the HOMER software (version 4.11).
Genome_build: mm9 (MGSCv37)
Supplementary_files_format_and_content: bedGraph

source_name: 3T3-L1 preadipocyte cell line

growth_protocol: Cells were maintained in DMEM-HG (high glucose), 10% CS until reaching confluent. 2 days before differentiation induction, the medium was replaced with fresh DMEM-HG, 10% CS. On day of differentiation induction (Day 0), the medium was replaced with DMEM-HG, 10% FBS supplemented with 1.67 uM insulin, 1 uM dexamethasone, and 115 ug/mL of IBMX. On Day 2, the medium was replaced with DMEM-HG, 10% FBS supplemented with 1.67 uM insulin. On Days 4 and 6, the medium was replaced with DMEM-HG, 10% FBS.

extract_protocol:
Genomic DNA was purified from cells using DNeasy Blood & Tissue Kit (QIAGEN) according to the manufacturer's protocol.
PCR-free whole-genome bisulfite-seq libraries were prepared with the tPBAT protocol as reported previously (Miura et al. 2019 Nucleic Acids Res, DOI: 10.1093/nar/gkz435).

treatment_protocol: In the first 2 days of differentiation, cells were cultured in the absence or presence of 100 uM deferoxamine.

value: preadipocyte cell line

value: DFO-treated

tag: time point

value: Day 8

tag: replicate

value: replicate 1


title: WGBS_Day8plus_rep3

value: replicate 3


title: WGBS_Day8minus_rep3

description:
Day8-noDFO.cpg.cover.bedGraph.gz
Day8-noDFO.cpg.methyl.bedGraph.gz

value: untreated


title: LF_Mock_rep3

city: Kitaadachi-gun

first: Hisanori

last: Takenobu

state: Saitama

address: 818, Komuro, Ina-machi

department: Research Institute for Clinical Oncology

institute: Saitama Cancer Center

description: human,  cell line

hyb_protocol: 600 ng of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 25 uL containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 25 uL of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4851C) for 17 hours at 65 °C in a rotating Agilent hybridization oven.  After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37 °C GE Wash buffer 2 (Agilent).

scan_protocol: Slides were scanned immediately after washing on the SureScan Agilent DNA Microarray Scanner (G4900DA) using one color scan setting for 8x60k array slides.

data_processing: The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.

source_name: Cranial neural crest cells from Li-Fraumeni syndrome patient-derived hiPSC, infected with Mock vector (pCDH-CMV-EF1a-puro),  Puromycin (1.0 μg/mL, 3 days)-selected bulk cells.

label_protocol: Cyanine-3 (Cy3) labeled cRNA was prepared from 200 ng RNA using the One-Color Low Input Quick Amp Labeling kit (Agilent) according to the manufacturer's instructions (Protocol version 6.9), followed by RNAeasy column purification (QIAGEN, Valencia, CA).  Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.

growth_protocol: LF cranial neural crest cells (cNCCs) were cultured in Iscove’s modified Dulbecco’s medium (Thermo Fisher Scientific)/Ham’s F-12 (Thermo Fisher Scientific) 1:1 supplemented with 1x chemically defined lipid concentrate (GIBCO), 15 mg/ml apo-transferrin (Sigma), 450 mM monothioglycerol (Sigma), 5 mg/ml purified BSA (Sigma), 7 mg/ml Insulin (FUJIFILM) and 50 μg/mL penicillin/streptomycin (Sigma). 20 ng/ml EGF (R&D) and 20 ng/ml FGF2 (FUJIFILM) were used to maintain cNCCs. Cultures were maintained at 37ºC under 5% CO2 in air.

extract_protocol: Total cellular RNA for preparing RT–PCR templates was extracted using ISOGEN II (Nippon Gene KK). RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was guaranteed by an RNA Integrity Number (RIN) > 8.

treatment_protocol: In the preparation of the overexpression/lentivirus, the packaging cell line HEK 293T (4 × 10^6) was plated on a 10-cm dish and co-transfected the next day with 1.5 μg of each of the plasmids (pCDH-CMV-EF1a-puro and pCDH-CMV-MYCN-EF1a-puro) and 2.0 μg of the packaging vectors (Sigma-Aldrich) using the Fugene HD transfection reagent (Promega) according to the manufacturer’s protocol. Medium was changed the next day and cells were cultured for another 24 h. Conditioned medium was collected and cleared of debris by filtering through a filter with 0.45-μm pores.  Thereafter, 1 × 105 cNCCs were seeded on each well of a 6-well plate, and infected by lentiviral-conditioned media with 4 μg/mL polybrene. The infected cNCCs were cultured in medium containing 1 μg/ml puromycin for 3 d, and were mixed with 0.3% agar dissolved in growth medium (CDMi + SB431542 + EGF + FGF2) and plated on top of a bottom layer of 0.5% agar dissolved in growth medium in a 35 mm dish. The transformed clones were isolated and injected into periadrenal adipose tissue of immune-deficient mice

value: LF cranial neural crest cells


title: LF_MYCN_CO-3_rep2

description: human,  osteosarcoma

source_name: Cranial neural crest cells from Li-Fraumeni syndrome patient-derived hiPSC, generated by injection into periadrenal adipose tissue of  immune-deficient mice.


title: daf-2, 60 Hours after eggs laid, replicate 2

city: Seoul

first: Daisy

middle: Sunghee

last: Lim

address: 1 Gwanak-ro, Gwanak-gu

country: South Korea

institute: Seoul National University

data_processing:
Kallisto v.0.46.1 was used to create cDNA index file and pseudoalignment (100 bootstraps)
Assembly: WBcel235, ensembl 103
Supplementary files format and content: tsv (tab-separated values) files generated by Kallisto, containing abundance information in estimated counts and tpm, for each sample

source_name: whole animal

growth_protocol: Synchronized daf-2 and daf-2; daf-42 embryos were obtained by placing 20 adult worms each on six NGM plates with OP50 for 2 hours at 15°C and then incubated at 25°C for dauer induction.

extract_protocol:
At 52 HAE and 60 HAE, the worms were harvested with M9 and washed with distilled water five times. The, 250 μL of TRIzol was added to 50 μL of washed worm harvest, and the worm samples were freeze-thawed 10–15 times for disruption. Harvest daf-2; daf-42 at 60 HAE included both live and dead L2d worms. RNA was extracted through chloroform—isopropanol precipitation.
Theragen Bio Inc. (South Korea) prepared a library with TruSeq Stranded mRNA Sample Prep Kit.

value: whole animal

value: worms were raised at 25 degree C


title: daf-2, 60 Hours after eggs laid, replicate 1


title: daf-2; daf-42, 52 Hours after eggs laid, replicate 3


title: daf-2, 52 Hours after eggs laid, replicate 2


title: daf-2; daf-42, 52 Hours after eggs laid, replicate 2


title: daf-2, 60 Hours after eggs laid, replicate 3


title: daf-2; daf-42, 60 Hours after eggs laid, replicate 3


title: LF_MYCN_CO-3_rep1

description: human,  osteosarcoma


title: daf-2; daf-42, 60 Hours after eggs laid, replicate 2


title: daf-2, 52 Hours after eggs laid, replicate 3


title: daf-2; daf-42, 52 Hours after eggs laid, replicate 1


title: LF_MYCN_CO-3_rep3

description: human,  osteosarcoma


title: LF_MYCN_TF-4_rep2

description: human,  cell line

source_name: Cranial neural crest cells from Li-Fraumeni syndrome patient-derived hiPSC, isolated from LF MYCN bulk cells in soft agar.


title: LF_Mock_rep2

description: human,  cell line


title: LF_Mock_rep1

description: human,  cell line


title: LF_MYCN_TF-4_rep3

description: human,  cell line


title: LF_MYCN_rep1

description: human,  cell line

source_name: Cranial neural crest cells from Li-Fraumeni syndrome patient-derived hiPSC, infected with MYCN overexpression vector (pCDH-CMV-MYCN-EF1a-puro),  Puromycin (1.0 μg/mL, 3 days)-selected bulk cells.


title: LF_MYCN_rep2

description: human,  cell line


title: daf-2; daf-42, 60 Hours after eggs laid, replicate 1


title: LF_MYCN_TF-4_rep1

description: human,  cell line


title: daf-2, 52 Hours after eggs laid, replicate 1


title: WGBS_Day8plus_rep2

value: replicate 2


title: WGBS_Day8minus_rep2


title: WGBS_Day8minus_rep1


title: 12B_M_998

city: La Jolla

first: Joseph

middle: R

last: Ecker

address: 10010 North Torrey Pines Road

department: Genomic Analysis Laboratory

institute: HHMI-Salk-Institute

data_processing:
Reads trimed with Cutadapt for adapters and low quality bases (Phred score >= 20)
The resulting files are mapped to in-silico bisulfite converted mm10 reference genome using Bismark.
The raw BAM file will be filtered using samtools to filter reads by MAPQ > 10 and use Picard to remove PCR duplicates
Tab-delimited files (ALLC files) describing the quantity of unmethylated and methylated cytosine base calls will be generated with in-house developing package using the filtered BAM files
Assembly: mm10
Supplementary files format and content: tab-delimited files (ALLC files) describing the quantity of unmethylated and methylated cytosine base calls. Columns are: 1. Chromosome, 2. Position, 3. Strand, 4. Cytosin Context, 5. methylated base call, 6. total mapped base call, 7. not used, place holder column
Library strategy: snmC-seq2

source_name: C57BL/6J Male P56 Mouse Brain

growth_protocol: All mice are C57BL/6J Male, brain dissected in P56

extract_protocol:
the brain of P56 male C57BL/6J mouse will be removed and cut into 600 microns thick coronal sections using a tissue matrix. Then each region will be microdissected from the coronal tissue sections based on identified landmarks and brain atlas coordinates, as defined in Allen brain atlas CCF3 (Avaliable at: http://atlas.brain-map.org). For each dissection region, the tissue from several littermate mice will be combined together into to separate replicate groups. The number of animals in a replicate depends on the size of the tissue. Nuclei preparation will be stained with the fluorescently labeled anti-NeuN body and nucleic acid stain Hoechst 33342.
Library prepared following protocol discribed in Luo, Chongyuan, Angeline Rivkin, Jingtian Zhou, Justin P. Sandoval, Laurie Kurihara, Jacinta Lucero, Rosa Castanon, et al. 2018. “Robust Single-Cell DNA Methylome Profiling with snmC-seq2.” Nature Communications 9 (1): 3824.

tag: brain region


title: GasN24_1

description:
GasN24_1_1.fq.gz
GasN24_1_2.fq.gz

value: muscle

value: female

value: 9 weeks

value: NRF


title: 12B_M_996


title: 12B_M_992


title: 12B_M_991


title: 12B_M_995


title: 12B_M_990


title: 12B_M_99


title: WGBS_Day2minus_rep2

description:
Day2-noDFO.cpg.cover.bedGraph.gz
Day2-noDFO.cpg.methyl.bedGraph.gz

tag: cell line

value: Day 2


title: WGBS_Day2plus_rep3

description:
Day2-DFO24h.cpg.cover.bedGraph.gz
Day2-DFO24h.cpg.methyl.bedGraph.gz

tag: cell type


title: WGBS_Day2minus_rep3

tag: treatment


title: WGBS_Day2plus_rep2


title: WGBS_Day2minus_rep1


title: WGBS_Day0_rep2

description:
Day0.cpg.cover.bedGraph.gz
Day0.cpg.methyl.bedGraph.gz

value: Day 0


title: 12B_M_983


title: 12B_M_986


title: WGBS_Day0_rep3


title: 12B_M_982


title: 12B_M_985


title: WGBS_Day0_rep1


title: 12B_M_984


title: WGBS_Day2plus_rep1


title: B1-HTR-8/SVneo-BHLHE40-ko rep1

city: Changsha

first: MI

middle: CHUN

last: MEI

state: HuNan

address: 138 Tongzipo Road, Yuelu District

department: Department of Obstetrics and Gynecology

institute: The Third Xiangya Hospital, Central South University

hyb_protocol: Samples were hybridized using Affymetrix hybridization kit materials • Heat cocktails at 99° for 5 minutes, then 42° for 5 minutes centrifuge at max speed for 1 minute (N.B. this deviates from Affy SOP). • Transfer 200μl of hyb solution to each array, then tape holes and parafilm. • Hybridize 16 hours at 45° at 60rpm• Fluidics washing is FS450_0001

scan_protocol: Affymetrix Gene ChIP Scanner 3000 7G

data_processing: Data were processed using GeneBASE software. MAT background correction were applied to probe intensities, gene level expression were summarized from the corrected intensities and then quantile normalized.

label_protocol: Samples were enzymatically fragmented and biotinylated using the WT Terminal Labeling Kit (Affymetrix)

extract_protocol: Trizol method was used to extract the total RNA of the samples. The total RNA was tested by nanodrop 2000 and Agilent Bioanalyzer 2100, and the qualified samples were put into the chip experiment.

treatment_protocol: HTR-8/SVneo Cells were cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, US) supplemented with 5% FBS, 100 U/ml of penicillin and 100 mg/ml of streptomycin (Invitrogen) at 37 °C in a 5% CO2 atmosphere.HTR-8/Svneo cells with BHLHE40 knockdown were cultured for 24 hours


title: A2-HTR-8/Svneo rep2

value: cultured for 24 hours


title: A1-HTR-8/Svneo rep3


title: B2-HTR-8/SVneo-BHLHE40-ko rep2

value: BHLHE40 knockdown were cultured for 24 hours


title: A1-HTR-8/Svneo rep1


title: 12B_M_981


title: 12B_M_979


title: B3-HTR-8/SVneo-BHLHE40-ko rep3


title: 12B_M_975


title: 12B_M_976


title: 12B_M_98


title: 12B_M_971


title: 12B_M_97


title: 12B_M_978


title: LF_MYCN_rep3

description: human,  cell line


title: 12B_M_964


title: 12B_M_974


title: 12B_M_969


title: 12B_M_961


title: 12B_M_96


title: 12B_M_959


title: 12B_M_960


title: 12B_M_957


title: 12B_M_967


title: 12B_M_955


title: 12B_M_968


title: 12B_M_958


title: 12B_M_954


title: 12B_M_952


title: 12B_M_953


title: 12B_M_951


title: 12B_M_95


title: 12B_M_980


title: 12B_M_949


title: 12B_M_947


title: 12B_M_948


title: 12B_M_946


title: 12B_M_940


title: 12B_M_941


title: 12B_M_942


title: 12B_M_94


title: 12B_M_938


title: 12B_M_939


title: 12B_M_936


title: 12B_M_937


title: 12B_M_933


title: 12B_M_935


title: 12B_M_931


title: 12B_M_929


title: 12B_M_956


title: 12B_M_93


title: 12B_M_930


title: 12B_M_927


title: 12B_M_934


title: 12B_M_928


title: 12B_M_926


title: 12B_M_924


title: 12B_M_950


title: 12B_M_922


title: 12B_M_920


title: 12B_M_92


title: 12B_M_945


title: 12B_M_918


title: 12B_M_919


title: 12B_M_916


title: 12B_M_917


title: 12B_M_915


title: 12B_M_910


title: 12B_M_914


title: 12B_M_91


title: 12B_M_909


title: 12B_M_905


title: 12B_M_908


title: 12B_M_904


title: 12B_M_901


title: 12B_M_907


title: 12B_M_902


title: 12B_M_90


title: 12B_M_898


title: 12B_M_9


title: 12B_M_899


title: 12B_M_925


title: 12B_M_897


title: 12B_M_894


title: 12B_M_923


title: 12B_M_895


title: 12B_M_896


title: 12B_M_891


title: 12B_M_890


title: 12B_M_89


title: 12B_M_881


title: 12B_M_882


title: 12B_M_912


title: 12B_M_885


title: 12B_M_883


title: 12B_M_88


title: 12B_M_879


title: 12B_M_878


title: 12B_M_873


title: 12B_M_872


title: 12B_M_877


title: 12B_M_874


title: 12B_M_87


title: 12B_M_868


title: 12B_M_865


title: 12B_M_871


title: 12B_M_870


title: 12B_M_867


title: 12B_M_864


title: 12B_M_863


title: 12B_M_862


title: 12B_M_861


title: 12B_M_86


title: 12B_M_892


title: 12B_M_859


title: 12B_M_858


title: 12B_M_889


title: 12B_M_880


title: 12B_M_857


title: 12B_M_854


title: 12B_M_851


title: 12B_M_852


title: 12B_M_855


title: 12B_M_850


title: 12B_M_875


title: 12B_M_849


title: 12B_M_869


title: 12B_M_847


title: 12B_M_846


title: 12B_M_840


title: 12B_M_845


title: 12B_M_841


title: 12B_M_84


title: 12B_M_834


title: 12B_M_838


title: 12B_M_836


title: 12B_M_837


title: 12B_M_835


title: 12B_M_832


title: 12B_M_830


title: 12B_M_83


title: 12B_M_829


title: 12B_M_827


title: 12B_M_828


title: 12B_M_856


title: 12B_M_826


title: 12B_M_825


title: 12B_M_824


title: 12B_M_823


title: 12B_M_821


title: 12B_M_820


title: 12B_M_82


title: 12B_M_85


title: 12B_M_819


title: 12B_M_822


title: 12B_M_816


title: 12B_M_812


title: 12B_M_811


title: 12B_M_833


title: 12B_M_817


title: 12B_M_810


title: 12B_M_81


title: 12B_M_809


title: 12B_M_814


title: 12B_M_807


title: 12B_M_806


title: 12B_M_802


title: 12B_M_804


title: 12B_M_800


title: 12B_M_80


title: 12B_M_805


title: 12B_M_797


title: 12B_M_796


title: 12B_M_798


title: 12B_M_793


title: 12B_M_795


title: 12B_M_791


title: 12B_M_790


title: 12B_M_792


title: 12B_M_789


title: 12B_M_788


title: 12B_M_787


title: 12B_M_786


title: 12B_M_815


title: 12B_M_783


title: 12B_M_782


title: 12B_M_781


title: 12B_M_779


title: 12B_M_784


title: 12B_M_78


title: 12B_M_778


title: 12B_M_775


title: 12B_M_773


title: 12B_M_774


title: 12B_M_772


title: 12B_M_771


title: 12B_M_77


title: 12B_M_769


title: 12B_M_768


title: 12B_M_794


title: 12B_M_764


title: 12B_M_767


title: 12B_M_765


title: 12B_M_763


title: 12B_M_762


title: 12B_M_76


title: 12B_M_759


title: 12B_M_758


title: 12B_M_785


title: 12B_M_780


title: 12B_M_756


title: 12B_M_754


title: 12B_M_757


title: 12B_M_755


title: 12B_M_753


title: 12B_M_750


title: 12B_M_748


title: 12B_M_751


title: 12B_M_75


title: 12B_M_747


title: 12B_M_744


title: 12B_M_746


title: 12B_M_749


title: 12B_M_743


title: 12B_M_745


title: 12B_M_770


title: 12B_M_742


title: 12B_M_740


title: 12B_M_738


title: 12B_M_739


title: 12B_M_741


title: 12B_M_737


title: 12B_M_74


title: 12B_M_736


title: 12B_M_760


title: 12B_M_734


title: 12B_M_732


title: 12B_M_731


title: 12B_M_73


title: 12B_M_726


title: 12B_M_728


title: 12B_M_723


title: 12B_M_721


title: 12B_M_729


title: 12B_M_722


title: 12B_M_720


title: 12B_M_717


title: 12B_M_719


title: 12B_M_71


title: 12B_M_710


title: 12B_M_714


title: 12B_M_709


title: 12B_M_708


title: 12B_M_707


title: 12B_M_705


title: 12B_M_703


title: 12B_M_704


title: 12B_M_72


title: 12B_M_702


title: 12B_M_701


title: 12B_M_700


title: 12B_M_7


title: 12B_M_699


title: 12B_M_70


title: 12B_M_698


title: 12B_M_695


title: 12B_M_697


title: 12B_M_696


title: 12B_M_725


title: 12B_M_712


title: 12B_M_692


title: 12B_M_690


title: 12B_M_713


title: 12B_M_69


title: 12B_M_689


title: 12B_M_688


title: 12B_M_687


title: 12B_M_684


title: 12B_M_686


title: 12B_M_683


title: 12B_M_682


title: 12B_M_691


title: 12B_M_681


title: 12B_M_68


title: 12B_M_678


title: 12B_M_673


title: 12B_M_674


title: 12B_M_675


title: 12B_M_672


title: 12B_M_671


title: 12B_M_670


title: 12B_M_67


title: 12B_M_669


title: 12B_M_663


title: 12B_M_665


title: 12B_M_694


title: 12B_M_664


title: 12B_M_693


title: 12B_M_660


title: 12B_M_661


title: 12B_M_659


title: 12B_M_662


title: 12B_M_66


title: 12B_M_658


title: 12B_M_656


title: 12B_M_653


title: 12B_M_677


title: 12B_M_655


title: 12B_M_651


title: 12B_M_650


title: 12B_M_652


title: 12B_M_649


title: 12B_M_643


title: 12B_M_644


title: 12B_M_64


title: 12B_M_642


title: 12B_M_646


title: 12B_M_637


title: 12B_M_635


title: 12B_M_634


title: 12B_M_633


title: 12B_M_632


title: 12B_M_639


title: 12B_M_641


title: 12B_M_631


title: 12B_M_628


title: 12B_M_657


title: 12B_M_626


title: 12B_M_630


title: 12B_M_63


title: 12B_M_625


title: 12B_M_624


title: 12B_M_62


title: 12B_M_65


title: 12B_M_619


title: 12B_M_621


title: 12B_M_616


title: 12B_M_618


title: 12B_M_645


title: 12B_M_623


title: 12B_M_613


title: 12B_M_612


title: 12B_M_614


title: 12B_M_610


title: 12B_M_609


title: 12B_M_611


title: 12B_M_608


title: 12B_M_607


title: 12B_M_603


title: 12B_M_604


title: 12B_M_605


title: 12B_M_60


title: 12B_M_629


title: 12B_M_6


title: 12B_M_596


title: 12B_M_599


title: 12B_M_598


title: 12B_M_594


title: 12B_M_595


title: 12B_M_597


title: 12B_M_59


title: 12B_M_590


title: 12B_M_588


title: 12B_M_583


title: 12B_M_584


title: 12B_M_580


title: 12B_M_58


title: 12B_M_577


title: 12B_M_581


title: 12B_M_574


title: 12B_M_576


title: 12B_M_572


title: 12B_M_570


title: 12B_M_57


title: 12B_M_568


title: 12B_M_589


title: 12B_M_565


title: 12B_M_566


title: 12B_M_567


title: 12B_M_562


title: 12B_M_575


title: 12B_M_561


title: 12B_M_592


title: 12B_M_559


title: 12B_M_558


title: 12B_M_556


title: 12B_M_554


title: 12B_M_553


title: 12B_M_587


title: 12B_M_551


title: 12B_M_550


title: 12B_M_549


title: 12B_M_548


title: 12B_M_546


title: 12B_M_545


title: 12B_M_543


title: 12B_M_542


title: 12B_M_54


title: 12B_M_539


title: 12B_M_538


title: 12B_M_537


title: 12B_M_535


title: 12B_M_533


title: 12B_M_536


title: 12B_M_563


title: 12B_M_600


title: 12B_M_560


title: 12B_M_530


title: 12B_M_53


title: 12B_M_529


title: 12B_M_528


title: 12B_M_525


title: 12B_M_524


title: 12B_M_523


title: 12B_M_522


title: 12B_M_521


title: 12B_M_520


title: 12B_M_55


title: 12B_M_519


title: 12B_M_52


title: 12B_M_515


title: 12B_M_513


title: 12B_M_511


title: 12B_M_510


title: 12B_M_51


title: 12B_M_506


title: 12B_M_509


title: 12B_M_508


title: 12B_M_505


title: 12B_M_503


title: 12B_M_502


title: 12B_M_504


title: 12B_M_499


title: 12B_M_5


title: 12B_M_498


title: 12B_M_497


title: 12B_M_50


title: 12B_M_501


title: 12B_M_495


title: 12B_M_494


title: 12B_M_492


title: 12B_M_490


title: 12B_M_489


title: 12B_M_491


title: 12B_M_517


title: 12B_M_518


title: 12B_M_516


title: 12B_M_488


title: 12B_M_487


title: 12B_M_514


title: 12B_M_485


title: 12B_M_481


title: 12B_M_482


title: 12B_M_480


title: 12B_M_484


title: 12B_M_479


title: 12B_M_478


title: 12B_M_477


title: 12B_M_474


title: 12B_M_475


title: 12B_M_476


title: 12B_M_48


title: 12B_M_470


title: 12B_M_473


title: 12B_M_47


title: 12B_M_469


title: 12B_M_466


title: 12B_M_465


title: 12B_M_463


title: 12B_M_462


title: 12B_M_460


title: 12B_M_579


title: 12B_M_461


title: 12B_M_49


title: 12B_M_458


title: 12B_M_456


title: 12B_M_454


title: 12B_M_455


title: 12B_M_486


title: 12B_M_483


title: 12B_M_450


title: 12B_M_45


title: 12B_M_453


title: 12B_M_447


title: 12B_M_449


title: 12B_M_446


title: 12B_M_443


title: 12B_M_442


title: 12B_M_445


title: 12B_M_44


title: 12B_M_437


title: 12B_M_438


title: 12B_M_436


title: 12B_M_434


title: 12B_M_431


title: 12B_M_435


title: 12B_M_433


title: 12B_M_430


title: 12B_M_428


title: 12B_M_427


title: 12B_M_429


title: 12B_M_457


title: 12B_M_451


title: 12B_M_425


title: 12B_M_424


title: 12B_M_423


title: 12B_M_422


title: 12B_M_444


title: 12B_M_421


title: 12B_M_420


title: 12B_M_418


title: 12B_M_42


title: 12B_M_416


title: 12B_M_415


title: 12B_M_414


title: 12B_M_413


title: 12B_M_412


title: 12B_M_410


title: 12B_M_411


title: 12B_M_408


title: 12B_M_409


title: 12B_M_403


title: 12B_M_404


title: 12B_M_402


title: 12B_M_405


title: 12B_M_400


title: 12B_M_40


title: 12B_M_399


title: 12B_M_4


title: 12B_M_398


title: 12B_M_394


title: 12B_M_393


title: 12B_M_395


title: 12B_M_392


title: 12B_M_388


title: 12B_M_384


title: 12B_M_385


title: 12B_M_386


title: 12B_M_383


title: 12B_M_419


title: 12B_M_379


title: 12B_M_38


title: 12B_M_378


title: 12B_M_376


title: 12B_M_374


title: 12B_M_377


title: 12B_M_372


title: 12B_M_407


title: 12B_M_370


title: 12B_M_369


title: 12B_M_371


title: 12B_M_373


title: 12B_M_368


title: 12B_M_365


title: 12B_M_364


title: 12B_M_360


title: 12B_M_396


title: 12B_M_361


title: 12B_M_36


title: 12B_M_359


title: 12B_M_389


title: 12B_M_357


title: 12B_M_358


title: 12B_M_355


title: 12B_M_356


title: 12B_M_354


title: 12B_M_352


title: 12B_M_353


title: 12B_M_351


title: 12B_M_35


title: 12B_M_382


title: 12B_M_375


title: 12B_M_347


title: 12B_M_348


title: 12B_M_345


title: 12B_M_344


title: 12B_M_346


title: 12B_M_343


title: 12B_M_367


title: 12B_M_340


title: 12B_M_34


title: 12B_M_335


title: 12B_M_363


title: 12B_M_339


title: 12B_M_337


title: 12B_M_333


title: 12B_M_334


title: 12B_M_342


title: 12B_M_33


title: 12B_M_329


title: 12B_M_328


title: 12B_M_324


title: 12B_M_341


title: 12B_M_323


title: 12B_M_322


title: 12B_M_320


title: 12B_M_349


title: 12B_M_32


title: 12B_M_319


title: 12B_M_327


title: 12B_M_350


title: 12B_M_330


title: 12B_M_317


title: 12B_M_315


title: 12B_M_318


title: 12B_M_313


title: 12B_M_312


title: 12B_M_31


title: 12B_M_338


title: 12B_M_309


title: 12B_M_308


title: 12B_M_306


title: 12B_M_331


title: 12B_M_310


title: 12B_M_311


title: 12B_M_303


title: 12B_M_301


title: 12B_M_304


title: 12B_M_305


title: 12B_M_302


title: 12B_M_300


title: 12B_M_30


title: 12B_M_298


title: 12B_M_297


title: 12B_M_299


title: 12B_M_295


title: 12B_M_293


title: 12B_M_296


title: 12B_M_292


title: 12B_M_314


title: 12B_M_288


title: 12B_M_290


title: 12B_M_287


title: 12B_M_286


title: 12B_M_284


title: 12B_M_283


title: 12B_M_281


title: 12B_M_282


title: 12B_M_279


title: 12B_M_28


title: 12B_M_277


title: 12B_M_278


title: 12B_M_274


title: 12B_M_269


title: 12B_M_275


title: 12B_M_276


title: 12B_M_267


title: 12B_M_3


title: 12B_M_266


title: 12B_M_265


title: 12B_M_263


title: 12B_M_262


title: 12B_M_260


title: 12B_M_291


title: 12B_M_255


title: 12B_M_254


title: 12B_M_258


title: 12B_M_252


title: 12B_M_26


title: 12B_M_294


title: 12B_M_251


title: 12B_M_249


title: 12B_M_248


title: 12B_M_280


title: 12B_M_243


title: 12B_M_241


title: 12B_M_239


title: 12B_M_244


title: 12B_M_246


title: 12B_M_25


title: 12B_M_24


title: 12B_M_240


title: 12B_M_245


title: 12B_M_247


title: 12B_M_235


title: 12B_M_234


title: 12B_M_232


title: 12B_M_231


title: 12B_M_230


title: 12B_M_23


title: 12B_M_227


title: 12B_M_224


title: 12B_M_222


title: 12B_M_223


title: 12B_M_225


title: 12B_M_221


title: 12B_M_220


title: 12B_M_253


title: 12B_M_219


title: 12B_M_217


title: 12B_M_213


title: 12B_M_212


title: 12B_M_214


title: 12B_M_207


title: 12B_M_208


title: 12B_M_206


title: 12B_M_205


title: 12B_M_237


title: 12B_M_203


title: 12B_M_204


title: 12B_M_229


title: 12B_M_20


title: 12B_M_211


title: 12B_M_200


title: 12B_M_199


title: 12B_M_198


title: 12B_M_197


title: 12B_M_2


title: 12B_M_194


title: 12B_M_190


title: 12B_M_192


title: 12B_M_189


title: 12B_M_187


title: 12B_M_188


title: 12B_M_19


title: 12B_M_186


title: 12B_M_201


title: 12B_M_183


title: 12B_M_185


title: 12B_M_215


title: 12B_M_180


title: 12B_M_18


title: 12B_M_177


title: 12B_M_179


title: 12B_M_181


title: 12B_M_178


title: 12B_M_174


title: 12B_M_193


title: 12B_M_191


title: 12B_M_173


title: 12B_M_175


title: 12B_M_172


title: 12B_M_170


title: 12B_M_166


title: 12B_M_169


title: 12B_M_167


title: 12B_M_164


title: 12B_M_165


title: 12B_M_163


title: 12B_M_160


title: 12B_M_161


title: 12B_M_159


title: 12B_M_21


title: 12B_M_162


title: 12B_M_158


title: 12B_M_157


title: 12B_M_155


title: 12B_M_153


title: 12B_M_1520


title: 12B_M_1516


title: 12B_M_1517


title: 12B_M_17


title: 12B_M_184


title: 12B_M_1514


title: 12B_M_1513


title: 12B_M_151


title: 12B_M_1509


title: 12B_M_1508


title: 12B_M_1512


title: 12B_M_154


title: 12B_M_1505


title: 12B_M_1507


title: 12B_M_1504


title: 12B_M_1501


title: 12B_M_1503


title: 12B_M_156


title: 12B_M_1499


title: 12B_M_1498


title: 12B_M_1494


title: 12B_M_1493


title: 12B_M_1492


title: 12B_M_1491


title: 12B_M_1490


title: 12B_M_149


title: 12B_M_1515


title: 12B_M_1486


title: 12B_M_1511


title: 12B_M_1488


title: 12B_M_1487


title: 12B_M_1485


title: 12B_M_1481


title: 12B_M_1480


title: 12B_M_1478


title: 12B_M_148


title: 12B_M_1479


title: 12B_M_150


title: 12B_M_1477


title: 12B_M_1476


title: 12B_M_1497


title: 12B_M_1472


title: 12B_M_1473


title: 12B_M_1475


title: 12B_M_1471


title: 12B_M_1465


title: 12B_M_1467


title: 12B_M_1466


title: 12B_M_1461


title: 12B_M_1489


title: 12B_M_1459


title: 12B_M_1462


title: 12B_M_1458


title: 12B_M_1457


title: 12B_M_1482


title: 12B_M_1454


title: 12B_M_1450


title: 12B_M_145


title: 12B_M_1455


title: 12B_M_1452


title: 12B_M_1449


title: 12B_M_1448


title: 12B_M_1447


title: 12B_M_1446


title: 12B_M_1444


title: 12B_M_1443


title: 12B_M_1442


title: 12B_M_1439


title: 12B_M_144


title: 12B_M_1438


title: 12B_M_1460


title: 12B_M_1436


title: 12B_M_1474


title: 12B_M_1434


title: 12B_M_1437


title: 12B_M_1430


title: 12B_M_1435


title: 12B_M_1429


title: 12B_M_1432


title: 12B_M_1428


title: 12B_M_1426


title: 12B_M_1424


title: 12B_M_1427


title: 12B_M_1422


title: 12B_M_142


title: 12B_M_1420


title: 12B_M_1419


title: 12B_M_1416


title: 12B_M_1417


title: 12B_M_1415


title: 12B_M_1418


title: 12B_M_1414


title: 12B_M_1413


title: 12B_M_141


title: 12B_M_1410


title: 12B_M_1412


title: 12B_M_1406


title: 12B_M_1404


title: 12B_M_1402


title: 12B_M_1400


title: 12B_M_1401


title: 12B_M_1395


title: 12B_M_1396


title: 12B_M_1398


title: 12B_M_1392


title: 12B_M_1394


title: 12B_M_1388


title: 12B_M_14


title: 12B_M_1389


title: 12B_M_1383


title: 12B_M_1382


title: 12B_M_1385


title: 12B_M_1381


title: 12B_M_1384


title: 12B_M_138


title: 12B_M_1377


title: 12B_M_1376


title: 12B_M_1375


title: 12B_M_1408


title: 12B_M_1403


title: 12B_M_1372


title: 12B_M_1371


title: 12B_M_1373


title: 12B_M_1370


title: 12B_M_137


title: 12B_M_1369


title: 12B_M_1368


title: 12B_M_1367


title: 12B_M_1386


title: 12B_M_1366


title: 12B_M_1365


title: 12B_M_1362


title: 12B_M_1360


title: 12B_M_1358


title: 12B_M_1357


title: 12B_M_1356


title: 12B_M_1380


title: 12B_M_1355


title: 12B_M_1354


title: 12B_M_1353


title: 12B_M_1351


title: 12B_M_1350


title: 12B_M_135


title: 12B_M_1348


title: 12B_M_1349


title: 12B_M_1361


title: 12B_M_1345


title: 12B_M_1343


title: 12B_M_1347


title: 12B_M_1342


title: 12B_M_1340


title: 12B_M_134


title: 12B_M_1363


title: 12B_M_1338


title: 12B_M_1337


title: 12B_M_1336


title: 12B_M_1359


title: 12B_M_1334


title: 12B_M_1333


title: 12B_M_1332


title: 12B_M_133


title: 12B_M_1330


title: 12B_M_1329


title: 12B_M_1328


title: 12B_M_1327


title: 12B_M_1324


title: 12B_M_1326


title: 12B_M_1321


title: 12B_M_1344


title: 12B_M_1320


title: 12B_M_132


title: 12B_M_1319


title: 12B_M_1318


title: 12B_M_1317


title: 12B_M_1316


title: 12B_M_1315


title: 12B_M_1339


title: 12B_M_1314


title: 12B_M_1335


title: 12B_M_1312


title: 12B_M_1313


title: 12B_M_131


title: 12B_M_1309


title: 12B_M_1306


title: 12B_M_1308


title: 12B_M_1305


title: 12B_M_1304


title: 12B_M_1302


title: 12B_M_1303


title: 12B_M_1300


title: 12B_M_130


title: 12B_M_13


title: 12B_M_1298


title: 12B_M_1295


title: 12B_M_1296


title: 12B_M_1294


title: 12B_M_1292


title: 12B_M_1290


title: 12B_M_1291


title: 12B_M_1285


title: 12B_M_1311


title: 12B_M_1281


title: 12B_M_1286


title: 12B_M_1282


title: 12B_M_1280


title: 12B_M_1323


title: 12B_M_1283


title: 12B_M_128


title: 12B_M_1278


title: 12B_M_1277


title: 12B_M_1301


title: 12B_M_1271


title: 12B_M_1274


title: 12B_M_1272


title: 12B_M_127


title: 12B_M_1269


title: 12B_M_1268


title: 12B_M_1265


title: 12B_M_1267


title: 12B_M_1266


title: 12B_M_1263


title: 12B_M_1288


title: 12B_M_126


title: 12B_M_1287


title: 12B_M_1258


title: 12B_M_1257


title: 12B_M_1251


title: 12B_M_1252


title: 12B_M_1256


title: 12B_M_1253


title: 12B_M_1250


title: 12B_M_125


title: 12B_M_1249


title: 12B_M_1279


title: 12B_M_1247


title: 12B_M_1246


title: 12B_M_1248


title: 12B_M_1243


title: 12B_M_1245


title: 12B_M_1241


title: 12B_M_1244


title: 12B_M_1234


title: 12B_M_1233


title: 12B_M_123


title: 12B_M_1239


title: 12B_M_124


title: 12B_M_1238


title: 12B_M_1341


title: 12B_M_1232


title: 12B_M_1237


title: 12B_M_1264


title: 12B_M_1231


title: 12B_M_1225


title: 12B_M_1229


title: 12B_M_1230


title: 12B_M_1223


title: 12B_M_1221


title: 12B_M_1222


title: 12B_M_1218


title: 12B_M_1224


title: 12B_M_1227


title: 12B_M_1226


title: 12B_M_1217


title: 12B_M_1214


title: 12B_M_1212


title: 12B_M_1216


title: 12B_M_1215


title: 12B_M_1240


title: 12B_M_1235


title: 12B_M_1211


title: 12B_M_1210


title: 12B_M_121


title: 12B_M_1209


title: 12B_M_1206


title: 12B_M_1205


title: 12B_M_1204


title: 12B_M_1203


title: 12B_M_1202


title: 12B_M_120


title: 12B_M_1201


title: 12B_M_12


title: 12B_M_1220


title: 12B_M_1197


title: 12B_M_1196


title: 12B_M_1195


title: 12B_M_1193


title: 12B_M_119


title: 12B_M_1352


title: 12B_M_1190


title: 12B_M_1191


title: 12B_M_1185


title: 12B_M_1188


title: 12B_M_1187


title: 12B_M_1182


title: 12B_M_1180


title: 12B_M_118


title: 12B_M_1179


title: 12B_M_1178


title: 12B_M_1207


title: 12B_M_1177


title: 12B_M_1174


title: 12B_M_1172


title: 12B_M_1173


title: 12B_M_1171


title: 12B_M_1170


title: 12B_M_1168


title: 12B_M_1199


title: 12B_M_1166


title: 12B_M_1167


title: 12B_M_1169


title: 12B_M_117


title: 12B_M_1165


title: 12B_M_1160


title: 12B_M_1161


title: 12B_M_1159


title: 12B_M_116


title: 12B_M_1158


title: 12B_M_1163


title: 12B_M_1157


title: 12B_M_1154


title: 12B_M_1153


title: 12B_M_1152


title: 12B_M_1150


title: 12B_M_115


title: 12B_M_1176


title: 12B_M_1149


title: 12B_M_1148


title: 12B_M_1147


title: 12B_M_1146


title: 12B_M_1145


title: 12B_M_1144


title: 12B_M_1143


title: 12B_M_1142


title: 12B_M_1198


title: 12B_M_1140


title: 12B_M_1141


title: 12B_M_1137


title: 12B_M_1162


title: 12B_M_1136


title: 12B_M_1135


title: 12B_M_1132


title: 12B_M_1134


title: 12B_M_1133


title: 12B_M_1156


title: 12B_M_1130


title: 12B_M_1128


title: 12B_M_1126


title: 12B_M_1125


title: 12B_M_1151


title: 12B_M_1122


title: 12B_M_1121


title: 12B_M_112


title: 12B_M_1118


title: 12B_M_1117


title: 12B_M_1115


title: 12B_M_1114


title: 12B_M_1112


title: 12B_M_1111


title: 12B_M_1110


title: 12B_M_111


title: 12B_M_1108


title: 12B_M_1109


title: 12B_M_1106


title: 12B_M_1107


title: 12B_M_1105


title: 12B_M_1104


title: 12B_M_1102


title: 12B_M_1100


title: 12B_M_1101


title: 12B_M_1129


title: 12B_M_1099


title: 12B_M_110


title: 12B_M_11


title: 12B_M_1098


title: 12B_M_1097


title: 12B_M_1095


title: 12B_M_1131


title: 12B_M_1119


title: 12B_M_1096


title: 12B_M_1094


title: 12B_M_1090


title: 12B_M_1092


title: 12B_M_1089


title: 12B_M_1088


title: 12B_M_1113


title: 12B_M_1085


title: 12B_M_1086


title: 12B_M_1087


title: 12B_M_1083


title: 12B_M_1081


title: 12B_M_1082


title: 12B_M_1084


title: 12B_M_1080


title: 12B_M_1079


title: 12B_M_1074


title: 12B_M_108


title: 12B_M_1075


title: 12B_M_1076


title: 12B_M_1077


title: 12B_M_1071


title: 12B_M_1073


title: 12B_M_1069


title: 12B_M_1070


title: 12B_M_1072


title: 12B_M_1066


title: 12B_M_107


title: 12B_M_1068


title: 12B_M_1067


title: 12B_M_1060


title: 12B_M_1065


title: 12B_M_1061


title: 12B_M_1058


title: 12B_M_1056


title: 12B_M_1057


title: 12B_M_1054


title: 12B_M_1053


title: 12B_M_1052


title: 12B_M_1051


title: 12B_M_1047


title: 12B_M_1049


title: 12B_M_1046


title: 12B_M_1044


title: 12B_M_1050


title: 12B_M_1041


title: 12B_M_1045


title: 12B_M_105


title: 12B_M_1043


title: 12B_M_1038


title: 12B_M_1035


title: 12B_M_1040


title: 12B_M_104


title: 12B_M_1036


title: 12B_M_1037


title: 12B_M_1064


title: 12B_M_1033


title: 12B_M_1031


title: 12B_M_106


title: 12B_M_1032


title: 12B_M_1055


title: 12B_M_1030


title: 12B_M_1027


title: 12B_M_1028


title: 12B_M_1024


title: 12B_M_1023


title: 12B_M_1022


title: 12B_M_1021


title: 12B_M_102


title: 12B_M_1019


title: 12B_M_1016


title: 12B_M_1018


title: 12B_M_1039


title: 12B_M_1015


title: 12B_M_1034


title: 12B_M_1012


title: 12B_M_101


title: 12B_M_1011


title: 12B_M_1014


title: 12B_M_1003


title: 12B_M_1005


title: 12B_M_1009


title: 12B_M_1004


title: 12B_M_1001


title: 12B_M_10


title: 12B_M_1000


title: 12A_M_3064


title: 12A_M_3062


title: 12A_M_3065


title: 12A_M_3063


title: 12A_M_3061


title: 12B_M_1020


title: 12A_M_3060


title: 12A_M_3058


title: 12A_M_3056


title: 12A_M_3054


title: 12A_M_3053


title: 12A_M_3050


title: 12A_M_3049


title: 12A_M_3051


title: 12B_M_1


title: 12A_M_3047


title: 12A_M_3048


title: 12A_M_3046


title: 12A_M_3044


title: 12A_M_3045


title: 12A_M_3043


title: 12B_M_1025


title: 12A_M_3041


title: 12A_M_3037


title: 12A_M_3036


title: 12A_M_3039


title: 12A_M_3038


title: 12A_M_3034


title: 12A_M_3057


title: 12A_M_3033


title: 12A_M_3035


title: 12A_M_3032


title: 12A_M_3052


title: 12A_M_3031


title: 12A_M_3030


title: 12A_M_3029


title: 12A_M_3027


title: 12A_M_3026


title: 12A_M_3025


title: 12A_M_3023


title: 12A_M_3022


title: 12A_M_3021


title: 12A_M_3019


title: 12A_M_3020


title: 12A_M_3018


title: 12A_M_3016


title: 12A_M_3017


title: 12A_M_3013


title: 12A_M_3014


title: 12A_M_3012


title: 12A_M_3011


title: 12A_M_3010


title: 12A_M_3008


title: 12A_M_3007


title: 12A_M_3042


title: 12A_M_3028


title: 12A_M_3024


title: 12A_M_3005


title: 12A_M_3006


title: 12A_M_3004


title: 12A_M_3002


title: 12A_M_3003


title: 12A_M_3001


title: 12A_M_2999


title: 12A_M_3059


title: 12A_M_3000


title: 12A_M_2998


title: 12A_M_2995


title: 12A_M_2997


title: 12A_M_3015


title: 12A_M_2994


title: 12A_M_2992


title: 12A_M_2991


title: 12A_M_2993


title: 12A_M_2989


title: 12A_M_2988


title: 12A_M_2987


title: 12A_M_2986


title: 12A_M_2983


title: 12A_M_2985


title: 12A_M_2982


title: 12A_M_2981


title: 12A_M_2979


title: 12A_M_2976


title: 12A_M_2996


title: 12A_M_2977


title: 12A_M_2975


title: 12A_M_2978


title: 12A_M_2974


title: 12A_M_2971


title: 12A_M_2973


title: 12A_M_2969


title: 12A_M_2967


title: 12A_M_2968


title: 12A_M_2965


title: 12A_M_2964


title: 12A_M_2963


title: 12A_M_2962


title: 12A_M_2961


title: 12A_M_2960


title: 12A_M_2980


title: 12A_M_2958


title: 12A_M_2956


title: 12A_M_2957


title: 12A_M_2955


title: 12A_M_2954


title: 12A_M_2953


title: 12B_M_113


title: 12A_M_2949


title: 12A_M_2951


title: 12A_M_2950


title: 12A_M_2947


title: 12A_M_2948


title: 12A_M_2970


title: 12A_M_2946


title: 12A_M_2945


title: 12A_M_2943


title: 12A_M_2942


title: 12A_M_2944


title: 12A_M_2941


title: 12A_M_2940


title: 12A_M_2938


title: 12A_M_2959


title: 12A_M_2933


title: 12A_M_2937


title: 12A_M_2935


title: 12A_M_2936


title: 12A_M_2934


title: 12A_M_2932


title: 12A_M_2952


title: 12A_M_2928


title: 12A_M_2930


title: 12A_M_2931


title: 12A_M_2929


title: 12A_M_2927


title: 12A_M_2924


title: 12A_M_2926


title: 12A_M_2925


title: 12A_M_2939


title: 12A_M_2923


title: 12A_M_2919


title: 12A_M_2922


title: 12A_M_2917


title: 12A_M_2921


title: 12A_M_2920


title: 12A_M_2914


title: 12A_M_2916


title: 12A_M_2913


title: 12A_M_2918


title: 12A_M_2910


title: 12A_M_2906


title: 12A_M_2912


title: 12A_M_2911


title: 12A_M_2904


title: 12A_M_2908


title: 12A_M_2909


title: 12A_M_2903


title: 12A_M_2907


title: 12A_M_2900


title: 12A_M_2901


title: 12A_M_2898


title: 12A_M_2899


title: 12A_M_2897


title: 12A_M_2896


title: 12A_M_2895


title: 12A_M_2894


title: 12A_M_2890


title: 12A_M_2888


title: 12A_M_3055


title: 12A_M_2892


title: 12A_M_2893


title: 12A_M_2889


title: 12A_M_2891


title: 12A_M_2915


title: 12A_M_2902


title: 12A_M_2885


title: 12A_M_2886


title: 12A_M_2884


title: 12A_M_2882


title: 12A_M_2883


title: 12A_M_3040


title: 12A_M_2880


title: 12A_M_2879


title: 12A_M_2877


title: 12A_M_2876


title: 12A_M_2905


title: 12A_M_2875


title: 12A_M_2871


title: 12A_M_2870


title: 12A_M_2869


title: 12A_M_2887


title: 12A_M_2872


title: 12A_M_2868


title: 12A_M_2874


title: 12A_M_2865


title: 12A_M_2864


title: 12A_M_2863


title: 12A_M_2862


title: 12A_M_2873


title: 12A_M_2867


title: 12A_M_2881


title: 12A_M_2859


title: 12A_M_2858


title: 12A_M_2860


title: 12A_M_2861


title: 12A_M_2857


title: 12A_M_2855


title: 12A_M_2854


title: 12A_M_2856


title: 12A_M_2850


title: 12A_M_2878


title: 12A_M_2853


title: 12A_M_2852


title: 12A_M_2851


title: 12A_M_2846


title: 12A_M_2845


title: 12A_M_2849


title: 12A_M_2843


title: 12A_M_2842


title: 12A_M_2848


title: 12A_M_2847


title: 12A_M_2841


title: 12A_M_2838


title: 12A_M_2840


title: 12A_M_2836


title: 12A_M_2827


title: 12A_M_2831


title: 12A_M_2828


title: 12A_M_2833


title: 12A_M_2829


title: 12A_M_2832


title: 12A_M_2826


title: 12A_M_2824


title: 12A_M_2844


title: 12A_M_2822


title: 12A_M_2821


title: 12A_M_2835


title: 12A_M_2818


title: 12A_M_2820


title: 12A_M_2816


title: 12A_M_2813


title: 12A_M_2814


title: 12A_M_2812


title: 12A_M_2811


title: 12A_M_999


title: 12A_M_998


title: 12A_M_2810


title: 12A_M_996


title: 12A_M_2866


title: 12A_M_992


title: 12A_M_2806


title: 12A_M_2804


title: 12A_M_2807


title: 12A_M_994


title: 12A_M_991


title: 12A_M_2803


title: 12A_M_995


title: 12A_M_2801


title: 12A_M_99


title: 12A_M_990


title: 12A_M_2800


title: 12A_M_987


title: 12A_M_2817


title: 12A_M_2815


title: 12A_M_2799


title: 12A_M_2798


title: 12A_M_2797


title: 12A_M_984


title: 12A_M_986


title: 12A_M_2796


title: 12A_M_982


title: 12A_M_2809


title: 12A_M_2794


title: 12A_M_981


title: 12A_M_2795


title: 12A_M_980


title: 12A_M_2793


title: 12A_M_2792


title: 12A_M_2790


title: 12A_M_977


title: 12A_M_976


title: 12A_M_2791


title: 12A_M_979


title: 12A_M_98


title: 12A_M_3009


title: 12A_M_2787


title: 12A_M_973


title: 12A_M_974


title: 12A_M_2786


title: 12A_M_972


title: 12A_M_2788


title: 12A_M_985


title: 12A_M_971


title: 12A_M_2784


title: 12A_M_97


title: 12A_M_969


title: 12A_M_2783


title: 12A_M_968


title: 12A_M_967


title: 12A_M_966


title: 12A_M_2780


title: 12A_M_965


title: 12A_M_2777


title: 12A_M_964


title: 12A_M_963


title: 12A_M_2776


title: 12A_M_975


title: 12A_M_2789


title: 12A_M_2775


title: 12A_M_2772


title: 12A_M_960


title: 12A_M_96


title: 12A_M_959


title: 12A_M_962


title: 12A_M_961


title: 12A_M_2768


title: 12A_M_2767


title: 12A_M_2770


title: 12A_M_2785


title: 12A_M_2774


title: 12A_M_958


title: 12A_M_2782


title: 12A_M_2769


title: 12A_M_957


title: 12A_M_2766


title: 12A_M_956


title: 12A_M_2765


title: 12A_M_2781


title: 12A_M_953


title: 12A_M_952


title: 12A_M_2764


title: 12A_M_2779


title: 12A_M_2763


title: 12A_M_2771


title: 12A_M_951


title: 12A_M_2762


title: 12A_M_2759


title: 12A_M_950


title: 12A_M_948


title: 12A_M_2761


title: 12A_M_2760


title: 12A_M_945


title: 12A_M_2758


title: 12A_M_946


title: 12A_M_2757


title: 12A_M_944


title: 12A_M_2756


title: 12A_M_955


title: 12A_M_954


title: 12A_M_2754


title: 12A_M_2755


title: 12A_M_943


title: 12A_M_2753


title: 12A_M_941


title: 12A_M_2751


title: 12A_M_2752


title: 12A_M_940


title: 12A_M_94


title: 12A_M_2749


title: 12A_M_939


title: 12A_M_2748


title: 12A_M_95


title: 12A_M_938


title: 12A_M_2746


title: 12A_M_2747


title: 12A_M_936


title: 12A_M_2745


title: 12A_M_947


title: 12A_M_937


title: 12A_M_2744