title: PDX003

city: ChangSha

first: Yongwei

last: Zhu

state: Hunan

address: No.87,  Xiangya Street

institute: CSU Xiangya hospital

data_processing:
Sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence, then mapped to GRCh38 whole genome using STAR (v2.7.5)with default parameters. BAM files of aligned reads were quantified by featureCounts (v2.0.1).
Fragments Per Kilobase of exon model per Million mapped fragments (FPKM) were calculated using a protocol from Trapnell C et al., Nat Biotechnol, 2010.
Assembly: GRCh38
Supplementary files format and content: tab-delimited text files include normalized read counts for each Sample

growth_protocol: The specimen was transported immediately from the operation room to the biological safety cabinet and washed 3 times with PBS (1X). We then transferred the washed tissue to 10cm2 dish to mince the tumor sample by using scalpel blades and surgical scissors. To obtain tumor pellets, the fragmented tumor tissue was moved to a 15 ml tube containing 0.05％ Trypsin-EDTA, pipetted for 7 minute and centrifuged at 1000 rpm for 5 min. Tumor pellets were cultured in serum-free NS cell self-renewal media (NS media) containing Neurocult NS-A Basal media (human), with 1X hormone mix (in house equivalent to N2), 2 mmol/L L-glutamine, 1X B27 supplement, 75 μg/mL BSA, 10 ng/mL bFGF and 10 ng/mL rhEGF, 2 μg/mL Heparin. After culturing for 3 to 10 days, GBM stem cells outgrow from the edge of the tumor pellets, while non-tumor cells were slowly removed after changing cell medium.

extract_protocol:
Total RNA was extracted from the tissues using Trizol (Invitrogen, Carlsbad, CA, USA) according to manual instruction.
mRNA libraries were prepared for sequencing using MGI DNA nanoball sequencing protocols.

treatment_protocol: We fabricated the polyacrylamide stiffness gel with various stiffness on glass coverslips according to our previous study.

value: glioma PDX


title: PDX004

status: Public on May 20 2023

type: SRA

country: China

library_source: transcriptomic

molecule: total RNA

organism: Homo sapiens

tag: tissue

tag: treatment


title: PDX002


title: Thatcher, Mock, 7 days, (Tc7), rep4

city: Ottawa

first: Michele

last: Loewen

state: Ontario

address: 100 Sussex Drive

country: Canada

institute: National Research Council of Canada (NRC)

data_processing:
Adaptor and quality trimming performed with fastp (Phred Score > 20, min read length > 50).
Quantification with Salmon in mapping-based mode
Differential gene expression analysis with DESeq2.
Assembly: IWGSC_RefSeqv2.1_HC_CDS_assembly
Supplementary files format and content: Transformed (vsd) transcript values from DESeq2 in tab-delimited format

organism: Triticum aestivum

source_name: flag leaf

growth_protocol: Plants were grown in Sunshine Mix #8 with a 16 hour/8 hour 22°C/17°C day/night cycle.

extract_protocol:
Total RNA was extracted using the Zymo Direct-zol RNA Miniprep kit (Infected samples) or the Monarch NEB Total RNA Miniprep kit (Mock samples)
cDNA libraries for infected samples were prepared using the TruSeq Stranded mRNA kit; cDNA libraries for mock samples were preparred using an Illumina Stranded mRNA, Ligation kit.

treatment_protocol: For infection, rust urediniospores were applied at the beginning of anthesis

value: flag leaf

tag: genotype

value: Thatcher

value: Mock 7 days


title: Thatcher, Mock, 14 days, (Tc14), rep1

value: Mock 14 days


title: Thatcher, Mock, 14 days, (Tc14), rep5


title: Thatcher, Mock, 7 days, (Tc7), rep2


title: Thatcher, Mock, 7 days, (Tc7), rep5


title: Thatcher, Mock, 14 days, (Tc14), rep2


title: Thatcher, Mock, 7 days, (Tc7), rep1


title: Thatcher, Mock, 7 days, (Tc7), rep3


title: PDX001


title: Day6, scRNAseq

city: Shanghai

first: junwei

last: shen

state: Shanghai

address: No.1239 Siping Road

institute: Tongji University

description: 10x Genomics

data_processing:
The demultiplexing, barcoded processing, gene counting and aggregation were made using the Cell Ranger software v2.1.0.
Assembly: GRCm38
Supplementary files format and content: Tab-separated values files and matrix files (barcodes.tsv, features.tsv, matrix.mtx).

organism: Mus musculus

source_name: entire heart

extract_protocol:
Heart samples from neonatal C57BL/6 mice were dissociated into single-cell suspensions. The tissues were minced with a razor blade, and then digested in digestion buffer with 0.75% Collagenase/Dispase. The cell suspension was filtered with cell strainers, and centrifuged. Subsequently, the cell pellet was resuspended with PBS and treated with 1X Red Blood Cell Lysis Solution to remove red blood cells. Trypan blue was used to check the viability of cells.
10x Library Preparation and Sequencing Beads with unique molecular identifiers (UMI) and cell barcodes are loaded to near saturation so that each cell is paired with a bead in a gel bead emulsion (GEM). After exposure to cell lysis buffer, polyadenylated RNA molecules hybridize to the beads. Beads were collected in one tube for reverse transcription. In cDNA synthesis, each cDNA molecule is labeled at the 5' end (ie, the 3' end of the messenger RNA transcript), and the UMI and cell tag indicate its cellular origin. Briefly, 10x beads are followed by second-strand cDNA synthesis, adapter ligation, and universal amplification. Sequencing libraries were prepared using randomly interrupted whole-transcriptome amplification products, enriched for 3' ends of transcripts linked to cellular barcodes and UMIs. All remaining steps, including library construction, were performed according to the standard manufacturer's protocol (CG000206 RevD, 10x Genomics). Sequencing libraries were quantified using the High Sensitivity DNA Chip (Agilent) and Qubit High Sensitivity DNA Assay (Thermo Fisher Scientific) on the Bioanalyzer 2100.

value: entire heart

tag: strain

tag: age

value: 6 days


title: Day4, scRNAseq

description: 10x Genomics

value: 4 days


title: PC9_shNC

city: Jinan

first: Jupeng

last: Yuan

state: Shandong

address: Jiyan road 440#

department: Cancer Research Center

institute: Shandong Cancer Hospital Affiliated to Shandong University

data_processing:
Raw data (raw reads) of fastq format were firstly processed through in-house perl scripts.
Index of the reference genome was built using Hisat2 v2.0.5 and paired-end clean reads were aligned to the reference genome using Hisat2 v2.0.5.
FeatureCounts v1.5.0-p3 was used to count the reads numbers mapped to each gene. And then FPKM of each gene was calculated based on the length of the gene and reads count mapped to this gene.
Genome_build: hg19
Supplementary_files_format_and_content: tab-delimited text files include gene count values for each Sample

molecule: polyA RNA

growth_protocol: Human PC9 cells were cultured in DMEM media supplied with 10% fetal bovine serum (Hyclone) at 37°C with 5% CO2.

extract_protocol:
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
A total amount of 3 μg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample.

treatment_protocol: Stable cell lines were obtained by transfecting with EPB41-depletion or control virus.

value: NSCLC cell line

tag: cell line

value: Transfected with control virus


title: Thatcher, Mock, 0 days, (Tc0), rep5

value: Mock 0 days


title: PC9_shEPB41-2

value: Transfected with EPB41-depletion virus


title: PC9_shEPB41-1


title: C643_PLVX

growth_protocol: Human C643 cells were cultured in DMEM media supplied with 10% fetal bovine serum (Hyclone) at 37°C with 5% CO2.

treatment_protocol: Stable cell lines were obtained by transfecting with SLC26A4-AS1 overexpressing or control virus.

value: thyroid cancer cell line


title: MDA-MB-468-sh-CONTROL_2

city: Ciudad de México

first: Mireya

last: Cisneros-Villanueva

state: Ciudad de Mexico

address: Periférico Sur No. 4809, Arenal Tepepan, 4809

country: Mexico

institute: Instituto Nacional de Medicina Genómica

data_processing:
The sequencing of RNA-seq libraries was carried out on the NextSeq 500 platform (Illumina, USA) at an average depth of 23 million raw paired reads per sample.
Quality control of raw fastq files was determined using FastQC (v. 0.11.9) and MultiQC (v. 1.11).
Adapter sequences, poor quality reads with Q values ≤22 and reads with fragment sizes ≤70 bp were filtered through Trimmomatic (v. 0.39).
The alignment of raw reads to GRCh38 (Homo sapiens) reference genome and the generation of bam files were carried out with Hisat2 (v. 2.1.0) and samtools, respectively.
Abundance estimation and mapped reads counting was performed with featureCounts (v. 2.0.0). All the analyses were carried out in the Unix-Bash environment of Linux platform
Assembly: hg38
Supplementary files format and content: tab-delimited text files include counts values for each Sample

source_name: cell line

growth_protocol: MDA-MB-468 cells were maintained in DMEM and HCC1187 cells were maintained in RPMI-1640 both were supplemented with 10% fetal bovine serum (FBS) in humidified atmosphere with 5% CO2 at 37C

extract_protocol:
RNA was harvested using TRIzol reagent (Invitrogen).  1 µg of total RNA was used for the construction of sequencing libraries
Paired-end RNA-seq libraries with fragments of 76 bp were generated from total RNA with the TruSeq Stranded mRNA Library Prep Kit (Illumina) following manufacturer's protocols

treatment_protocol: Cells were transfected during 48 hours

value: cell line

tag: cell type

value: basal-like cells

value: shCONTROL

value: sh-LacZ rep2


title: Day2, scRNAseq

description: 10x Genomics

value: 2 days


title: C643_AS1

value: Transfected with SLC26A4-AS1 overexpressing virus


title: Thatcher, Mock, 0 days, (Tc0), rep4

value: flag leaf

value: Thatcher


title: Thatcher, Mock, 0 days, (Tc0), rep3


title: Thatcher, Mock, 0 days, (Tc0), rep2


title: Day1, scRNAseq

description: 10x Genomics

value: 1 day


title: Thatcher, Mock, 0 days, (Tc0), rep1


title: Thatcher, Infected, 14 days, (Tc+LR14), rep5

value: Infected 14 days


title: Thatcher, Infected, 14 days, (Tc+LR14), rep4


title: T20-N: Adjacent tissues of No.20 patient

data_processing:
Raw data (raw reads) of fastq format were firstly processed through in-house perl scripts.
Use Stringtie software to compare the assembled samples with reference transcriptomes
after assembly, combine the assembled transcripts
then use FEELnc software to perform lncRNA identification on the combined transcripts; except for lncRNA, the others are mRNA.
Genome_build: hg19
Supplementary_files_format_and_content: tab-delimited text files include gene count values for each Sample

source_name: Adjacent tissues

extract_protocol:
Total RNA was extracted with TRIZOL reagents following the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA).
A total amount of 3 μg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample.

tag: disease state

value: thyroid cancer (THC)

tag: gender

value: Female

tag: ct

tag: cn


title: T20-T: Tumor tissues of No.20 patient

source_name: Tumor tissues


title: T16-T: Tumor tissues of No.16 patient


title: T16-N: Adjacent tissues of No.16 patient


title: Thatcher, Infected, 14 days, (Tc+LR14), rep2


title: T16-LN: Metastatic lymph nodes of No.16 patient

source_name: Metastatic lymph nodes


title: Thatcher, Infected, 14 days, (Tc+LR14), rep3


title: T15-LN: Metastatic lymph nodes of No.15 patient


title: T15-T: Tumor tissues of No.15 patient


title: Thatcher, Infected, 14 days, (Tc+LR14), rep1


title: T13-LN: Metastatic lymph nodes of No.13 patient

value: Male


title: Thatcher, Infected, 7 days, (Tc+LR7), rep5

value: Infected 7 days


title: T15-N: Adjacent tissues of No.15 patient


title: T13-T: Tumor tissues of No.13 patient


title: Thatcher, Infected, 7 days, (Tc+LR7), rep4


title: T13-N: Adjacent tissues of No.13 patient


title: HCC1187-sh-CONTROL_1

value: sh-LacZ rep1


title: heat_41_3

city: Changsha

first: Lan

last: Zhang

state: --- Select One ---

address: 172 Tongzipo road, Yuelu District, Changsha city, Hunan Province

institute: 中南大学

data_processing:
Trimmomatic  v0.36
Raw data (raw reads) were processed using Trimmomatic
The reads containing ploy-N and the low quality reads were removed to obtain the clean reads.
The clean reads were mapped to reference genome using hisat2.
Assembly: NCBI_GRCh38.p13
Supplementary files format and content: tab-delimited text files include RPKM values for each Sample
Supplementary files format and content: tab-delimited text files include counts values for each Sample

source_name: skin

extract_protocol:
Total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion) following the manufacturer’s protocol.
The libraries were constructed using TruSeq Stranded mRNA LTSample Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer’s instructions.

value: skin

value: HaCaT

value: human immortalized keratinocyte

value: not applicable

value: 41℃ for 3 days, 1 hour per day


title: heat_41_2


title: Thatcher, Infected, 7 days, (Tc+LR7), rep3


title: T20-LN: Metastatic lymph nodes of No.20 patient


title: CC2

city: Nanjing

first: Yu

last: Fan

state: Jiangsu

address: Xianlin 138 Road

institute: Nanjing University of Chinese Medicine

description: C57BL/6J male mice,Cocaine, Day78

data_processing:
Illumina Casava2.19 software used for basecalling.
Sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence, then mapped to hg38 whole genome using hisat2 v2.0.5 with parameters hisat2 [options]* -x <hisat2-idx> {-1 <m1> -2 <m2> | -U <r> | --sra-acc <SRA accession number>} [-S <hit>]
Fragments Per Kilobase of exon per Megabase of library size (FPKM) were calculated using a protocol from Trapnell,Cole et al., Transcript assembly and quantification by RNA-seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat. Biotechnol, 2010. In short, exons from all isoforms of a gene were merged to create one meta-transcript. The number of reads falling in the exons of this meta-transcript were counted and normalized by the size of the meta-transcript and by the size of the library.
Assembly: hg38
Supplementary files format and content: tab-delimited text files include FPKM values for each Sample ...

extract_protocol:
Acute extraction of mouse brain tissue within 30-60 minutes after CPP test, flash frozen on dry ice, extracting PFC brain tissue and RNA was harvested using Trizol reagent. Illumina TruSeq RNA Sample Prep Kit (Cat#FC-122-1001) was used with 1 ug of total RNA for the construction of sequencing libraries.
RNA libraries were prepared for sequencing using standard Illumina protocols

value: mesenchymal stem cells

value: WT

value: P28-P42inject 15 mg/kg cocaine,P75 pre-test,P76-P77inject 1 mg/kg cocaine before CPP training,P78 test and tissue collection

tag: time

value: Day 5


title: CC4

description: C57BL/6J male mice,Cocaine, Day78


title: SC6

description: C57BL/6J male mice,Control, Day78

value: P28-P42inject saline,P75 pre-test,P76-P77inject 1 mg/kg cocaine before CPP training,P78 test and tissue collection


title: CC5

description: C57BL/6J male mice,Cocaine, Day78

value: Day 0


title: CC6

description: C57BL/6J male mice,Cocaine, Day78


title: CC1

description: C57BL/6J male mice,Cocaine, Day78


title: CC3

description: C57BL/6J male mice,Cocaine, Day78


title: SC5

description: C57BL/6J male mice,Control, Day78


title: heat_41_1


title: Thatcher, Infected, 7 days, (Tc+LR7), rep1

value: flag leaf

value: Thatcher


title: Thatcher, Infected, 7 days, (Tc+LR7), rep2


title: heat_37_3

value: 37℃ for 3 days, 1 hour per day


title: SC4

description: C57BL/6J male mice,Control, Day78


title: T8-LN: Metastatic lymph nodes of No.8 patient

value: thyroid cancer (THC)

value: Female


title: heat_37_2

value: Human immortalized keratinocyte


title: T8-N: Adjacent tissues of No.8 patient


title: SC3

description: C57BL/6J male mice,Control, Day78


title: heat_37_1

value: skin


title: SC2

description: C57BL/6J male mice,Control, Day78


title: T7-T: Tumor tissues of No.7 patient

value: Male


title: 6d-2

city: Guiyang; Huaxi

first: dan

middle: dan

last: Li

address: Jiaxiunan road

institute: Guizhou university

description: seed

data_processing:
The libraries were sequenced on an llumina Novaseq 6000 platform and 150 bp paired-end reads  were generated. About xxxxx raw reads for each sample were generated. Raw reads of fastq format  were firstly processed using fastp  1 and the low quality reads were removed to obtain the clean reads. Then about xxxxxx clean reads for each sample were retained for subsequent analyses.
The clean reads were mapped to the reference genome using HISAT2 2. FPKM3 of each gene was calculated and the read counts of each gene were obtained by HTSeq-count4 . PCA analysis were performed using R (v 3.2.0) to evaluate the biological duplication of samples.
Hierarchical cluster analysis of DEGs was performed using R (v 3.2.0) to demonstrate the  expression pattern of genes in different groups and samples. The radar map of top 30 genes was  drew to show the expression of up-regulated or down-regulated DEGs using R packet ggradar .
Gene Set Enrichment Analysis (GSEA) was performed using GSEA software8-9. The analysis was used a predefined gene set, and the genes were ranked according to the degree of differential expression in the two types of samples. Then it is tested whether the predefined gene set was enriched at the top or bottom of the ranking list.
Assembly: https://safflower.scuec.edu.cn/download.html
Supplementary files format and content: Txt file including  FPKM value in each sample

organism: Carthamus tinctorius

source_name: seed

extract_protocol:
Total RNA was extracted using the TRIzol reagent (Invitrogen, CA, USA) according to the  manufacturer’s protocol. RNA purity and quantification were evaluated using the NanoDrop 2000  spectrophotometer (Thermo Scientific, USA). RNA integrity was assessed using the Agilent 2100  Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA).
Then the libraries were constructed using  VAHTS Universal V6 RNA-seq Library Prep Kit according to the manufacturer’s instructions. The  transcriptome sequencing and analysis were conducted by OE Biotech Co., Ltd. (Shanghai, China).

value: seed


title: SC1

description: C57BL/6J male mice,Control, Day78


title: 6d-3

description: seed


title: 6d-1

description: seed


title: C4_3

city: hangzhou

first: jiafeng

last: chen

state: zhejiang

address: yuhangtang road 866

institute: zhejiang university

data_processing:
Reads obtained from the sequencing machines includes raw reads containing adapters or low quality bases which will affect the following assembly and analysis. Thus, to get high quality clean reads, reads were further filtered by Cutadapt (https://cutadapt.readthedocs.io/en/stable/,version:cutadapt-1.9)
We aligned reads of all samples to the research species reference genome using HISAT2 (https://daehwankimlab.github.io/hisat2/,version:hisat2-2.0.4) package, which initially remove a portion of the reads based on quality information accompanying each read and then maps the reads to the reference genome.
The mapped reads of each sample were assembled using StringTie (http://ccb.jhu.edu/software/stringtie/,version:stringtie-1.3.4d) with default parameters. Then, all transcriptomes from all samples were merged to reconstruct a comprehensive transcriptome using gffcompare software(http://ccb.jhu.edu/software/stringtie/gffcompare.shtml,version:gffcompare-0.9.8). After the final transcriptome was generated, StringTie and ballgown (http://www.bioconductor.org/packages/release/bioc/html/ballgown.html) were used to estimate the expression levels of all transcripts and perform expression abundance for mRNAs by calculating FPKM (fragment per kilobase of transcript per million mapped reads) value.
Genes differential expression analysis was performed by DESeq2 software between two different groups (and by edgeR between two samples). The genes with the parameter of false discovery rate (FDR) below 0.05 and absolute fold change ≥ 2 were considered differentially expressed genes. Differentially expressed genes were then subjected to enrichment analysis of GO functions and KEGG pathways.
Assembly: not provided

organism: Danio rerio

source_name: AB

growth_protocol: raised and maintained at 28°C on 14 h light/10 h dark rhythmical cycles in the zebrafish core facility of Zhejiang University School of Medicine.

extract_protocol:
Total RNA was extracted using Trizol reagent following the manufacturer's procedure. The total RNA quantity and purity were analysis of Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit, high-quality RNA samples with RIN number > 7.0 were used to construct sequencing library.
After total RNA was extracted, mRNA was purified from total RNA (5ug) using Dynabeads Oligo (dT) with two rounds of purification. Following purification, the mRNA was fragmented into short fragments using divalent cations under elevated temperature (Magnesium RNA Fragmentation Module under 94℃ 5-7min). Then the cleaved RNA fragments were reverse-transcribed to create the cDNA by SuperScript™ II Reverse Transcriptase, which were next used to synthesise U-labeled second-stranded DNAs with E. coli DNA polymerase I, RNase H and dUTP Solution. An A-base was then added to the blunt ends of each strand, preparing them for ligation to the indexed adapters. Each adapter contained a T-base overhang for ligating the adapter to the A-tailed fragmented DNA. Dual-index adapters were ligated to the fragments, and size selection was performed with AMPureXP beads. After the heat-labile UDG enzyme treatment of the U-labeled second-stranded DNAs, the ligated products were amplified with PCR by the following conditions: initial denaturation at 95℃ for 3 min; 8 cycles of denaturation at 98℃ for 15 sec, annealing at 60℃ for 15 sec, and extension at 72℃ for 30 sec; and then final extension at 72℃ for 5 min. The average insert size for the final cDNA librarys were 300±50 bp. At last, we performed the 2×150bp paired-end sequencing (PE150) on an Illumina Novaseq™ 6000 (LC-Bio Technology CO., Ltd., Hangzhou, China) following the vendor's recommended protocol.

treatment_protocol: Zebrafish embryos were subjected to control or BPC treatments at 4 hours post fertilization

value: AB

value: BPC 4μM

value: 72 hpf


title: T7-N: Adjacent tissues of No.7 patient


title: C4_1


title: T6-LN: Metastatic lymph nodes of No.6 patient


title: FLOWER3

description: flower

source_name: flower

value: flower


title: T6-T: Tumor tissues of No.6 patient


title: T8-T: Tumor tissues of No.8 patient


title: FLOWER2

description: flower


title: DMSO_3

value: DMSO


title: Immune cells, HDM group, mouse 3

city: New Brunswick

first: Qi

last: Yang

state: NJ

address: 89 French St

country: USA

institute: Rutgers RWJMS

description: 10x Genomics

library_source: transcriptomic single cell

data_processing:
Library was generated using chromium 5' gene expression kit for mesenchymal cekks and using chromiun 3' CellMultiplex kit for immune cells(10Xgenomincs).
Reference genome mouse (mm10) was used for read alignment and gene expression quantification using Cellranger v3.1.0 pipelines.
The Cellranger pipelines include Cellranger mkfastq, Cellranger count, Cellranger Aggr, and Cellranger reanalyze pipelines.
These pipelines combine Chromium specific algorithms with the RNA-seq aligner STAR.
Assembly: mm10
Supplementary files format and content: mtx and tsv for processed data, bam for each sample

source_name: Lung

extract_protocol:
Mesenchymal cells and immune cells from the lungs were sorted based on their surface marker expressions. Total RNA extracted following the instructions of the 10X genomics kit
Mesenchymal cells scRNA-seq libraries were prepared using the 5’ single cell gene expression kit and immune cells scRNA-seq libraries were prepared using 3' CellMultiplex kit (10x Genomics) accoding to the manufacturer’s instruction.

treatment_protocol: Mice were injected with PBS or 25ug HDM (i.n.) on day 1, 8, 15, 16 and 17. Then on day 18, we euthanized the mice and collect mesenchymal cells and immune cells from the lungs.

value: Immune cells

value: Lung

value: 6-8 weeks old

value: House dust mite (HDM) challenged


title: FLOWER1

description: flower


title: T5-LN: Metastatic lymph nodes of No.5 patient


title: Immune cells, HDM group, mouse 1

description: 10x Genomics


city: Daejeon

first: Su-Jin

last: Baek

address: 1672, Yuseong-daero

country: South Korea

institute: KOREA INSTITUTE of ORIENTAL MEDICINE

description:
iPSCs treated with 10 μg/mL ECR (replicate 3)
N10_H_3

data_processing:
Quality of raw sequence data was checked via the FastQC and adapter or low quality bases were trimmed with TrimGalore
After trimming, the reads were aligned to the human reference genome (hg38) using the STAR aligner
Read count and transcripts per million (TPM) were calculated as gene expression abundance using the RSEM software (v1.3.3) and gene annotation GRCh38.84 Gene Transfer Format (GTF) file
Assembly: hg38
Supplementary files format and content: tab-delimited text files include count values for each Sample

source_name: iPSC

extract_protocol:
One mg of total RNA was processed for preparing mRNA sequencing library using MGIEasy RNA Directional Library Prep Kit (MGI Tech, China) according to manufacturer’s instruction.
Library was quantified by QauntiFluor ssDNA System (Promega, WI, USA).

value: iPSC

value: 10 ug/mL ECR


description:
iPSCs treated with 10 μg/mL ECR (replicate 2)
N10_H_2


title: LEAF3

description: leaf

source_name: leaf

value: leaf


title: LEAF2

description: leaf


title: Immune cells, PBS group, mouse 4

description: 10x Genomics

value: PBS


title: Immune cells, PBS group, mouse 3

description: 10x Genomics


title: Immune cells, HDM group, mouse 2

description: 10x Genomics


title: LEAF1

description: leaf


title: T7-LN: Metastatic lymph nodes of No.7 patient

value: thyroid cancer (THC)


title: Immune cells, HDM group, mouse 4.

description: 10x Genomics


description:
iPSCs treated with 5 μg/mL ECR (replicate 3)
N10_L_3

value: 5 ug/mL ECR


title: Immune cells, PBS group, mouse 2

description: 10x Genomics


title: STEM3

description: stem

source_name: stem

value: stem


title: T4-LN: Metastatic lymph nodes of No.4 patient

value: Female


title: T5-N: Adjacent tissues of No.5 patient

value: Male


title: Immune cells, PBS group, mouse 1

description: 10x Genomics


title: DMSO_1

value: AB


title: Mesenchymal cells, HDM group, sample 2, pooled from 3 mice

description: 10x Genomics

value: Mesenchymal cells


description:
iPSCs treated with 5 μg/mL ECR (replicate 1)
N10_L_1


title: T6-N: Adjacent tissues of No.6 patient


title: STEM1

description: stem


description:
iPSCs treated with vehicle (replicate 3)
control_3

value: vehicle


title: T3-LN: Metastatic lymph nodes of No.3 patient


title: Mesenchymal cells, HDM group, sample 1, pooled from 3 mice

description: 10x Genomics


title: ROOT3

description: root

source_name: root

value: root


description:
iPSCs treated with vehicle (replicate 2)
control_2


title: T5-T: Tumor tissues of No.5 patient


title: ROOT2

description: root


title: Mesenchymal cells, PBS group, sample 2, pooled from 3 mice

description: 10x Genomics


title: T3-T: Tumor tissues of No.3 patient


description:
iPSCs treated with 10 μg/mL ECR (replicate 1)
N10_H_1


description:
iPSCs treated with vehicle (replicate 1)
control_1


title: ROOT1

description: root


title: PLC-shYAP1-RNA-seq

status: Public on May 19 2023

city: Guangzhou

first: Linchong

last: Sun

state: Guangdong

address: B2 Building, South China University of Technology, No. 382, Waihuandong Road, Guangzhou Higher Education Mega Center, Panyu District

institute: South China University of China

description: total RNA

data_processing:
As to RIP-seq and RNA-seq data, raw data (raw reads) of fastq format were firstly processed through in-house perl scripts.
Reads were alligned to the human genome hg19 with BOWTIE2 v2.2.6 and clean reads were aligned to the reference genome using TopHat2 v2.1.0.
Cuffdiff v2.2.1 was used to count the reads numbers mapped to each gene.
Genome_build: hg19

source_name: Human PLC cells

growth_protocol: Human HEK293T cells and PLC cells were cultured in DMEM (GIBCO) with 10% FBS. Cells were maintained in a water-jacketed incubator with 5% CO2 in 37 °C.

extract_protocol:
As to RIP-seq and RNA-seq, RNA was extracted using TRIZOL Reagent (Life technologies) following the manufacturer’s instructions and checked for a RIN number to inspect RNA integrity by an Agilent Bioanalyzer 2100.Qualified total RNA was further purified by RNeasy micro kit and RNase-Free DNase Set. Sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample.
RNA-sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample.
RIP-seq and RNA-seq

value: Human-derived PLC cells

tag: rip-antibody

value: Human liver


title: PLC-shENO1-RNA-seq

description: total RNA


title: PLC-NTC-RNA-seq

description: total RNA


title: T3-N: Adjacent tissues of No.3 patient


title: STEM2

description: stem


description:
iPSCs treated with 5 μg/mL ECR (replicate 2)
N10_L_2


title: T4-T: Tumor tissues of No.4 patient


title: T4-N: Adjacent tissues of No.4 patient


title: Mesenchymal cells, PBS group, sample 1, pooled from 3 mice

description: 10x Genomics

value: Lung

value: 6-8 weeks old


title: atoh1b-MO_V

city: Dresden

first: Stefan

last: Hans

address: Fetscherstraße 105

department: CRTD - Center for Regenerative Therapies Dresden

country: Germany

institute: Technische Universität Dresden

data_processing:
bcl2fastq2: bcl2fastq2 2.19.1
alignment: Reads were aligned with gsnap (v2017-11-15) to the GRCz10 genome and the support of Ensembl version 87 gene annotation to detect splice sites (gsnap.sse42 -d GRCz10 --gunzip -A sam -t 14 --use-sarray=1 --input-buffer-size=500000 --output-buffer-size=500000 -B 5 -N 1 -n 1 -s EnsemblGene-87.ss.GRCz10.iit)
gene counts: Uniquely mapped reads were assigned to gene with the tool featureCounts (v1.5.3) (featureCounts -a EnsemblGene-87.GRCz10.TR.gtf -s 2 --primary -Q 1 -T 8
Assembly: GRCz10
Supplementary files format and content: bfx870.counts.tsv.gz is a tab separated file from featureCounts. Each row is a gene. Each columns is a gene information (Start, End, Strand, …) or a sample. Each cell is the reads per gene per sample.

source_name: pax8:DsRed-positive cells

extract_protocol:
RNA isolation from sorted cells was performed using the Total RNA purification Micro Kit (Norgen Biotek) following the manufacturer’s protocol.
mRNA was isolated from 30 ng total RNA by poly-dT enrichment using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) according to the manufacturer’s instructions. Samples were then directly subjected to the workflow for strand-specific RNA-Seq library preparation (Ultra II Directional RNA Library Prep, NEB). For ligation custom adaptors were used (Adaptor-Oligo 1: 5'-ACA CTC TTT CCC TAC ACG ACG CTC TTC CGA TCT-3', Adaptor-Oligo 2: 5'-P-GAT CGG AAG AGC ACA CGT CTG AAC TCC AGT CAC-3'). After ligation, adapters were depleted by an XP bead purification (Beckman Coulter) adding the beads solution in a ratio of 1:0.9. Dual unique indexing was done during the following PCR enrichment (14 cycles) using custom amplification primers carrying the same sequence for i7 and i5 index (Primer 1: AAT GAT ACG GCG ACC ACC GAG ATC TAC AC NNNNNNNN ACA TCT TTC CCT ACA CGA CGC TCT TCC GAT CT, Primer 2: CAA GCA GAA GAC GGC ATA CGA GAT NNNNNNNN GTG ACT GGA GTT CAG ACG TGT GCT CTT CCG ATC T). After two more XP bead purifications (1:0.9), libraries were quantified using the Fragment Analyzer (Agilent). For Illumina flowcell production, samples were equimolarly pooled and sequenced 75bp single read on an Illumina HiSeq 2500.

treatment_protocol: Zebrafish embryos (wt or atoh1b-MO samples) were removed from their chorions by pronase treatment, followed by deyolking at 4°C in 0.5 % Ginzburg-Ringer without CaCl2. Dissociation was conducted in trypsin-EDTA on ice. When embryos were completely dissociated, the reaction was stopped by adding Hi-FBS. The cells were pelleted, washed with PBS, resuspended in PBS and passed through a 40 μM mesh filter prior to cell sorting. FACS was performed using an Aria II cell sorter (BD Biosciences). Forward and side scatter were used to gate for live, single cells, out of which DsRed-positive cells were sorted and collected.

value: pax8:DsRed-positive cells

value: OEPD and nephric anlagen

value: dlx3b/4b knockdown

tag: selection

value: FAC-sorted


title: atoh1b-MO_IV


title: atoh1b-MO_II


title: ctrl_II

value: wt


title: ctrl_III


title: HEK293T cells-input RNA

description: total RNA

source_name: Human HEK293T cells

value: Human-derived HEK293T cells

value: human embryonic kidney


title: wt_2

data_processing:
bcl2fastq2: bcl2fastq2 2.17.1.14
alignment: Reads were aligned with gsnap (v2017-06-20) to the GRCz10 genome and the support of Ensembl version 87 gene annotation to detect splice sites (gsnap.sse42 -d GRCz10 --gunzip -A sam -t 14 --use-sarray=1 --input-buffer-size=500000 --output-buffer-size=500000 -B 5 -N 1 -n 1 -s EnsemblGene-87.ss.GRCz10.iit)
gene counts: Uniquely mapped reads were assigned to gene with the tool featureCounts (v1.5.2) (featureCounts -a EnsemblGene-87.GRCz10.TR.gtf -s 2 --primary -Q 1 -T 8
Assembly: GRCz10
Supplementary files format and content: bfx784.counts.tsv.gz is a tab separated file from featureCounts. Each row is a gene. Each columns is a gene information (Start, End, Strand, …) or a sample. Each cell is the reads per gene per sample.

extract_protocol:
RNA isolation from sorted cells was performed using the Total RNA purification Micro Kit (Norgen Biotek) following the manufacturer’s protocol.
mRNA was isolated from 25 ng total RNA by poly-dT enrichment using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) according to the manufacturer’s instructions. Samples were then directly subjected to the workflow for strand-specific RNA-Seq library preparation (Ultra II Directional RNA Library Prep, NEB). For ligation custom adaptors were used (Adaptor-Oligo 1: 5'-ACA CTC TTT CCC TAC ACG ACG CTC TTC CGA TCT-3', Adaptor-Oligo 2: 5'-P-GAT CGG AAG AGC ACA CGT CTG AAC TCC AGT CAC-3'). After ligation, adapters were depleted by an XP bead purification (Beckman Coulter) adding the beads solution in a ratio of 1:0.9. Dual unique indexing was done during the following PCR enrichment (14 cycles) using custom amplification primers carrying the same sequence for i7 and i5 index (Primer 1: AAT GAT ACG GCG ACC ACC GAG ATC TAC AC NNNNNNNN ACA TCT TTC CCT ACA CGA CGC TCT TCC GAT CT, Primer 2: CAA GCA GAA GAC GGC ATA CGA GAT NNNNNNNN GTG ACT GGA GTT CAG ACG TGT GCT CTT CCG ATC T). After two more XP bead purifications (1:0.9), libraries were quantified using the Fragment Analyzer (Agilent). For Illumina flowcell production, samples were equimolarly pooled and sequenced 75bp single read on an Illumina HiSeq 2500.

treatment_protocol: Zebrafish embryos (wt or dlx3b/4b-MO samples) were removed from their chorions by pronase treatment, followed by deyolking at 4°C in 0.5 % Ginzburg-Ringer without CaCl2. Dissociation was conducted in trypsin-EDTA on ice. When embryos were completely dissociated, the reaction was stopped by adding Hi-FBS. The cells were pelleted, washed with PBS, resuspended in PBS and passed through a 40 μM mesh filter prior to cell sorting. FACS was performed using an Aria II cell sorter (BD Biosciences). Forward and side scatter were used to gate for live, single cells, out of which DsRed-positive cells were sorted and collected.


title: HEK293T cells-ENO1-RIP-seq

description: Immunoprecipitated RNA with ENO1 antibody


title: ctrl_I


title: dlx3p4_MO_2


title: dlx3p4_MO_1


title: wt_3


title: wt_1


title: ST3neg_5_HTO

city: CAMBRIDGE

first: Duncan

middle: Matthew

last: Morgan

state: MA

address: 500 Main St

department: Koch Institute

institute: MIT

data_processing:
Cell hashing data was aligned to HTO barcodes using CITE-seq-Count v1.4.2.
Raw read processing of scRNA-seq reads was performed as previously described35. Briefly, reads were aligned to the hg38 reference genome or Mmul_10 reference genome and collapsed by cell barcode and unique molecular identifier (UMI). Whole transcriptome data was further analyzed in Seurat66. First, cells with less than 300 unique genes detected and genes detected in fewer than 5 cells were filtered out. Variable features were then determined using the FindVariableFeatures function, and the ScaleData function was then used to regress out the number of RNA features in each cell. The number of principal components used for visualization was determined by examination of the elbow plot, and two-dimensional embeddings were generated using uniform manifold approximation and projection (UMAP). Clusters were determined using Louvain clustering, as implemented in the FindClusters function in Seurat. For analysis of ST3-reactive B cell data, clusters of cells that exhibited high expression of mitochondrial genes or markers associated with other cell phenotypes (i.e. monocytes) were moved and the data was reprocessed. Differential gene expression analysis was performed for each cluster and between indicated cell populations using the FindMarkers function.
Processing of BCR sequence data was performed with pRESTO and Change-O from the Immcantation software suite69,70. First, potential errors in cell barcode and UMI sequence were corrected for errors up to one nucleotide mismatch with a directional UMI collapse, as implemented in UMI-Tools71. Data was filtered by Q-score using the FilterSeq.py function to remove any sequences with an average Q score below 25. Then, the MaskPrimers.py function was used to annotate sequences with the correct isotype (R1 index) and V-region (R2) primer, as well as to mask the corresponding primer regions. The PairSeq.py function was used to retain only read pairs that passed both the FilterSeq.py and MaskPrimers.py processing steps. The data was then segregated by cell barcode and UMI, and the BuildConsensus.py function was used to call consensus sequences for read 1 index and read 2 of each barcode and UMI separately. The AssemblePairs.py function was used to assemble these consensus sequences into a single overlapping sequence. The resulting sequences were then analyzed with IgBlast72 using reference sequences provided by IMGT.
Assembly: hg38
Assembly: mmul_10
Supplementary files format and content: tab separated values files and comma separated files

organism: Macaca mulatta

source_name: PBMC

extract_protocol:
Whole human blood was obtained from Research Blood Components, LLC (Watertown, MA), and PBMC were isolated by means of density gradient centrifugation and cryopreserved. Upon use, PBMC were thawed into Aim-V medium (ThermoFisher). B cells were isolated using an EasySep Human Pan-B Cell Enrichment kit (Stemcell Technologies, Vancouver, Canada).
Biotinylated pneumococcal serotype 3 polysaccharide with a 5% biotin load (ST3, from Pfizer EPBD) was coupled with high concentration streptavidin-phycoerythrin (SA-PE) and streptavidin-allophycocyanin (SA-APC, both from BioLegend) in separate reactions. Biotinylated ST3 and the fluorophore-conjugated streptavidin reagents were prepared with PBS at 4X working solutions of 20 µg/mL and 200 µg/mL, respectively. Equal volumes of ST3 and SA-PE; and, separately, equal volumes of ST3 and SA-APC working solutions were combined, thoroughly mixed and incubated at 4 ºC protected from light for a minimum incubation period of 60 minutes with thorough mixing at 30 minutes and directly prior to use. Coupled ST3-PE and ST3-APC solutions were maintained at 4 ºC, protected from light until use.
Seq-Well

value: PBMC

value: B cell


title: KMB_rep2a

city: Hong Kong

first: Xudong

last: Liu

address: Tat Chee Avenue Kowloon Tong

institute: City University of Hong Kong

description:
KMB_rep2
The group contained kidney marrow with blood (KMB).

library_source: genomic

data_processing:
All the raw Nanopore data (fast5 type) were basecalled by Bonito v.0.6.1 (https://github.com/nanoporetech/bonito) with the dna_r9.4.1_e8_sup@3.3 model and the argument of “--modified_bases 5mc” for 5mC to .bam files with methylation information. To get the site-level modification proportion, modbam2bed v.0.6.3 (https://github.com/epi2me-labs/modbam2bed) was used to profile the bam file. To obtain the proportion of covered CpG sites in ONT methods, only the site that had at least one forward and one reverse sequencing read coverage was counted. To assess the methylation status of each chromosome region, the chromosome was divided into 100Kb fragments. The 5Kb fragments upstream of the transcription start site (TSS) were defined as the promoter region. The regional methylation proportion was then calculated for each fragment using an in-house script, which can be found at https://github.com/lxd98/zebrafish_scripts.
Assembly: zebrafish GRCz11
Supplementary files format and content: The content of .bed files including the methylation propotion at difference regional level.

molecule: genomic DNA

source_name: kidney marrow with blood

extract_protocol:
Wild-type Tubingen (TU) zebrafish lines were obtained from the Zebrafish International Resource Center (ZIRC, USA). The study received approval from the Committee on the Use of Laboratory and Research Animals (CULATR) at the University of Hong Kong (HKU). Kidney marrow from 8-month-old WT zebrafish was collected, dissociated in 0.9X PBS via pipetting, and filtered through a 40 µm nylon cell strainer (Corning, NY, USA). Some samples underwent treatment with eBioscience™ 1x RBC Lysis Buffer (Invitrogen, MA, USA) to lyse red blood cells, and these samples were designated as the KM group. Samples without red blood cell lysis were categorized as the KMB group. The resulting cells were centrifuged at 500g for 5 minutes, resuspended, filtered, and washed twice with 0.9X PBS for DNA extraction. Genomic DNA was extracted using QIAamp DNA Kits (Qiagen, Germany).
Approximately 2 μg of Genomic DNA extracted from the kidney marrow cells of zebrafish was used to prepare the library with a ligation sequencing kit (LSK-110, ONT). The libraries were sequenced on MinION Mk1B after loading to R9.4.1 flow cells (FLO-MIN106D, ONT).

value: kidney marrow with blood


title: KMB_rep2c


title: KMB_rep2b


title: KM_rep2a

description:
KM_rep2
The group contained kidney marrow without blood (KM).

source_name: kidney marrow

value: kidney marrow


title: KM_rep1b

description:
KM_rep1
The group contained kidney marrow without blood (KM).


title: KM_rep2b


title: KMB_rep1a

description:
KMB_rep1
The group contained kidney marrow with blood (KMB).


title: KM_rep2c


title: KM_rep1c


title: ST3neg_5_LC


title: KMB_rep1b


title: KMB_rep1c


title: dlx3p4_MO_3


title: ST3neg_5_HC


title: KM_rep1a


title: ST3neg_5_DGE


title: ST3neg_4_HTO


title: ST3neg_4_LC


title: ST3neg_4_DGE


title: ST3neg_4_HC


title: ST3neg_3_HTO


title: ST3neg_3_LC


title: ST3neg_3_HC


title: ST3neg_3_DGE


title: ST3pos_4_HC


title: ST3pos_4_HTO


title: ST3pos_3_LC


title: ST3pos_4_LC


title: ST3pos_4_DGE


title: ST3pos_3_HC


title: ST3neg_2_LC


title: ST3neg_1_LC


title: ST3pos_2_LC


title: ST3neg_1_DGE


title: ST3neg_2_HTO


title: ST3pos_3_HTO


title: ST3pos_3_DGE


title: ST3pos_2_DGE


title: ST3pos_2_HTO


title: ST3pos_2_HC


title: ST3pos_1_HTO


title: ST3neg_1_HC


title: ST3pos_1_LC


title: ST3pos_1_HC


title: ST3pos_1_DGE


title: ST3neg_2_HC


title: ST3neg_1_HTO


title: ST3neg_2_DGE


title: MEF, CFAE+8 medium plus compounds (MCE Cat.No. HY-L009,RackCode: SA00129416)

city: 海淀区

first: Jun

state: 北京市

address: 北京市海淀区颐和园路5号北京大学王克桢楼207

institute: Peking University

data_processing:
DRUG-seq2 data was processed to obtain expression matrix of gene counts according to drop-seq analysis protocol (https://github.com/broadinstitute/Drop-seq). Briefly, reads for each well were split through 10-base well and 10-base plate barcodes and then mapped to the mouse transcriptome with the mm10 reference index and Ensembl gene references by STAR (Dobin et al., 2012). The unique alignments were further demultiplexed based on the transcript UMI for each gene. The UMIs of each gene were counted as its raw counts. CPM (counts of exon model per million mapped reads) matrix was transferred from the raw count matrix and used for hierarchical clustering, Pearson correlation analysis, and PCA analysis.
Assembly: mm10
Supplementary files format and content: Tab-separated values files and count matrix files

source_name: embryo

extract_protocol: Samples with compound treatments in a 96 well-plate were washed with pre-warmed PBS. 60 μL lysis buffer containing 50 mM Tris-HCl pH8.0, 75 mM KCl, 6% Ficoll PM-400, 0.15% TritonX-100, and 0.5U/μL RNaseOUT (RNaseOut should be added before use) were added to each well. The 96-well plate was then incubated on a shaker at 1,000 rpm for 15 minutes. The lysates were then transferred to a new 96-well PCR plate.10 μL reverse transcription mix containing 40 mM MgCl2, 4 μM strand switching primer (TSO), 0.1 mM dNTP, 8 U/μL Maxima reverse transcriptase, 0.4 μL 1:100 diluted ERCC, and RNaseOUT (RNaseOut should be added before use) were added to each well, mixed, and lightly centrifuged for several seconds to collect all reactants at the bottom of well. A 96-well PCR plate was then incubated at 42°C for 2 hours with a hot lid of 105°C. After RT-PCR, products from a 96-well plate were pooled into a clean 50 mL tube and then twice purified according to the manufacturer’s instructions of DNA clean & concentrator-100 kit (ZYMO Research, D4029) and Agecourt RNA Clean XP beads (BECKMAN COULTER, A63987), respectively. The purified products were dissolved in 32 μL H2O. Next, 4 μL exonuclease I (NEB, M0293S) and 4 μL exonuclease I buffer were added to the above solution and then incubated in 37°C for 30 min, followed by 85°C for 15 min to digest the residual primers. Products from the digestion were directly amplified with Kapa HIFI PCR ReadyMix (Roche, KK2602) and 1 μM DRUG-seq2 pre-amplification primer via the PCR cycling conditions: 96°C for 1 min, 5 cycles of 98°C for 20 s, 58°C for 4 min, and 72°C for 6 min; 10 cycles of 98°C for 20 s, 60°C for 30 s, and 72°C for 6 min, with a final extension at 72°C for 10 min. PCR products were purified and collected with 25 μL H2O according to the manufacturer’s instructions (Agecourt RNA Clean XP beads; BECKMAN COULTER, A63987). Pre-amplification products were quantified via Qubit 4.0 (Thermofisher, Q33240) and then analysed on an Agilent Fragment Analyzer 5400. The result of fragment distribution should be a single peak at around 1,500~3,000bp with slight degradation. Qualified samples were then processed to obtain the pre-library that included fragmentation with Tn5 and 3’-end enrichment with 5 μM DRUG-seq2_P5_PCR primer and 5 μM Plastech--seq indexing primer (TruePrepTM DNA Library Prep Kit V2 for Illumina (Vazyme, TD502). The PCR procedure included: 72°C for 3 min, 98°C for 30 s, 15 cycles of 98°C for 15s, 60°C for 30s, and 72°C for 3 min, with a final extension of 72°C for 5 min. The pre-library was used to select 250-450 bp products with Agencourt AMPure XP beads (BECKMAN COULTER, A63880). Briefly, 50 μL pre-library was transferred to a 1.5 mL tube and mixed with 35 μL AMPure XP beads until the solution appeared homogeneous. After incubating for 5 min at room temperature, the 1.5 mL tube was placed on the magnet stand for 5 min until the solution was clear and the beads were collected at one corner of the tube. The supernatant was aspirated and discarded. The beads were washed twice with 500 μL fresh 80% ethanol for 30s at room temperature. During the washing, the beads remained on the magnet strand to avoid disturbing the separated magnetic beads. The tube containing the magnetic beads was then air-dried for 5 min with the cap open. After that, the washed and air-dried beads were eluted by 26.5 μL nuclease free water. The tube was incubated at room temperature for 5 min and then placed on the magnet stand for 5 min to separate the beads and solution. Finally, 25 μl solution was collected into a new 1.5 mL tube without disturbing the beads. This solution (library) was quantified via Qubit 4.0 (Thermofisher, Q33240), and then analysed on the Agilent Fragment Analyzer 5400. The qualified library was sequenced on the Illumina Hiseq X-ten platform.

value: embryo

value: embryonic fibroblast

value: WT

value: CFAE+8 medium plus compounds (MCE Cat.No. HY-L009,RackCode:SA00129416) for 4 days


title: MEF, CFAE+8 medium plus compounds (MCE Cat.No. HY-L009,RackCode: 50101921)

address: 北京市海淀区颐和园路5号北京大学王克桢楼207

value: CFAE+8 medium plus compounds (MCE Cat.No. HY-L009,RackCode:50101921) for 4 days


title: 12H2_M_1531

city: La Jolla

first: Joseph

middle: R

last: Ecker

state: CA

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-seq3

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

value: MRN


title: 12H2_M_1533


title: 12H2_M_1524


title: 12H2_M_1530


title: 12H2_M_1527


title: 12H2_M_1532


title: 12H2_M_1534


title: 12H2_M_1535


title: 12H2_M_1529


title: 12H2_M_1523


title: 12H2_M_1525


title: 12H2_M_1520


title: 12H2_M_1521


title: 12H2_M_1518


title: 12H2_M_1519


title: 12H2_M_1515


title: 12H2_M_1514


title: 12H2_M_1516


title: 12H2_M_1512


title: 12H2_M_1517


title: 12H2_M_1513


title: MEF, CFAE+8 medium plus compounds (MCE Cat.No. HY-L009,RackCode: SA00153143)

address: 北京市海淀区颐和园路5号北京大学王克桢楼207

value: CFAE+8 medium plus compounds (MCE Cat.No. HY-L009,RackCode:SA00153143) for 4 days


title: 12H2_M_1528


title: 12H2_M_1511


title: 12H2_M_1510


title: 12H2_M_1526


title: 12H2_M_1522


title: 12H2_M_1509


title: 12H2_M_1508


title: 12H2_M_1506


title: 12H2_M_1503


title: 12H2_M_1505


title: 12H2_M_1504


title: 12H2_M_1499


title: 12H2_M_1497


title: 12H2_M_1500


title: 12H2_M_1502


title: 12H2_M_1487


title: 12H2_M_1492


title: 12H2_M_1485


title: 12H2_M_1493


title: 12H2_M_1491


title: 12H2_M_1490


title: 12H2_M_1488


title: 12H2_M_1486


title: 12H2_M_1494


title: 12H2_M_1498


title: 12H2_M_1496


title: 12H2_M_1489


title: 12H2_M_1484


title: 12H2_M_1483


title: 12H2_M_1482


title: 12H2_M_1507


title: 12H2_M_1479


title: 12H2_M_1480


title: 12H2_M_1501


title: 12H2_M_1495


title: 12H2_M_1481


title: 12H2_M_1478


title: 12H2_M_1477


title: 12H2_M_1476


title: 12H2_M_1475


title: 12H2_M_1472


title: 12H2_M_1469


title: 12H2_M_1474


title: 12H2_M_1473


title: 12H2_M_1468


title: 12H2_M_1465


title: 12H2_M_1460


title: 12H2_M_1467


title: 12H2_M_1464


title: 12H2_M_1462


title: 12H2_M_1459


title: 12H2_M_1471


title: 12H2_M_1458


title: 12H2_M_1466


title: 12H2_M_1461


title: 12H2_M_1454


title: 12H2_M_1457


title: 12H2_M_1456


title: 12H2_M_1455


title: 12H2_M_1470


title: 12H2_M_1463


title: 12H2_M_1450


title: 12H2_M_1451


title: 12H2_M_1446


title: 12H2_M_1448


title: 12H2_M_1447


title: 12H2_M_1453


title: 12H2_M_1449


title: 12H2_M_1439


title: 12H2_M_1445


title: 12H2_M_1440


title: 12H2_M_1442


title: 12H2_M_1441


title: 12H2_M_1443


title: 12H2_M_1423


title: 12H2_M_1424


title: 12H2_M_1430


title: 12H2_M_1434


title: 12H2_M_1436


title: 12H2_M_1435


title: 12H2_M_1444


title: 12H2_M_1438


title: 12H2_M_1432


title: 12H2_M_1425


title: 12H2_M_1431


title: 12H2_M_1437


title: 12H2_M_1428


title: 12H2_M_1429


title: 12H2_M_1427


title: 12H2_M_1426


title: 12H2_M_1433


title: 12H2_M_1418


title: 12H2_M_1421


title: 12H2_M_1417


title: 12H2_M_1416


title: 12H2_M_1419


title: 12H2_M_1422


title: 12H2_M_1420


title: 12H2_M_1413


title: 12H2_M_1415


title: 12H2_M_1414


title: 12H2_M_1409


title: 12H2_M_1410


title: 12H2_M_1411


title: 12H2_M_1408


title: 12H2_M_1404


title: 12H2_M_1407


title: 12H2_M_1401


title: 12H2_M_1399


title: 12H2_M_1403


title: 12H2_M_1400


title: 12H2_M_1406


title: 12H2_M_1405


title: 12H2_M_1397


title: 12H2_M_1393


title: 12H2_M_1394


title: 12H2_M_1402


title: 12H2_M_1398


title: 12H2_M_1396


title: 12H2_M_1395


title: 12H2_M_1390


title: 12H2_M_1391


title: 12H2_M_1392


title: 12H2_M_1388


title: 12H2_M_1386


title: 12H2_M_1389


title: 12H2_M_1385


title: 12H2_M_1412


title: 12H2_M_1384


title: 12H2_M_1383


title: 12H2_M_1382


title: 12H2_M_1377


title: 12H2_M_1373


title: 12H2_M_1381


title: 12H2_M_1364


title: 12H2_M_1370


title: 12H2_M_1361


title: 12H2_M_1363


title: 12H2_M_1378


title: 12H2_M_1369


title: 12H2_M_1362


title: 12H2_M_1380


title: 12H2_M_1371


title: 12H2_M_1374


title: 12H2_M_1368


title: 12H2_M_1375


title: 12H2_M_1379


title: 12H2_M_1366


title: 12H2_M_1365


title: 12H2_M_1376


title: 12H2_M_1372


title: 12H2_M_1367


title: 12H2_M_1359


title: 12H2_M_1357


title: 12H2_M_1358


title: 12H2_M_1360


title: 12H2_M_1355


title: 12H2_M_1354


title: 12H2_M_1356


title: 12H2_M_1352


title: 12H2_M_1353


title: 12H2_M_1278


title: 12H2_M_1275


title: 12H2_M_1271


title: 12H2_M_1270


title: 12H2_M_1276


title: 12H2_M_1277


title: 12H2_M_1279


title: 12H2_M_1273


title: 12H2_M_1351


title: 12H2_M_1350


title: 12H2_M_1265


title: 12H2_M_1263


title: 12H2_M_1347


title: 12H2_M_1267


title: 12H2_M_1268


title: 12H2_M_1264


title: 12H2_M_1345


title: 12H2_M_1344


title: 12H2_M_1274


title: 12H2_M_1269


title: 12H2_M_1346


title: 12H2_M_1348


title: 12H2_M_1349


title: 12H2_M_1266


title: 12H2_M_1262


title: 12H2_M_1261


title: 12H2_M_1260


title: 12H2_M_1272


title: 12H2_M_1259


title: 12H2_M_1258


title: 12H2_M_1257


title: 12H2_M_1256


title: 12H2_M_1255


title: 12H2_M_1238


title: 12H2_M_1254


title: 12H2_M_1251


title: 12H2_M_1243


title: 12H2_M_1244


title: 12H2_M_1240


title: 12H2_M_1253


title: 12H2_M_1248


title: 12H2_M_1246


title: 12H2_M_1252


title: 12H2_M_1239


title: 12H2_M_1235


title: 12H2_M_1230


title: 12H2_M_1237


title: 12H2_M_1231


title: 12H2_M_1232


title: 12H2_M_1245


title: 12H2_M_1234


title: 12H2_M_1250


title: 12H2_M_1241


title: 12H2_M_1233


title: 12H2_M_1242


title: 12H2_M_1249


title: 12H2_M_1236


title: 12H2_M_1247


title: 12H2_M_1227


title: 12H2_M_1228


title: 12H2_M_1229


title: 12H2_M_1226


title: 12H2_M_1225


title: 12H2_M_1223


title: 12H2_M_1221


title: 12H2_M_1219


title: 12H2_M_1217


title: 12H2_M_1220


title: 12H2_M_1218


title: 12H2_M_1216


title: 12H2_M_957


title: 12H2_M_1222


title: 12H2_M_952


title: 12H2_M_950


title: 12H2_M_956


title: 12H2_M_955


title: 12H2_M_951


title: 12H2_M_949


title: 12H2_M_948


title: 12H2_M_947


title: 12H2_M_954


title: 12H2_M_943


title: 12H2_M_944


title: 12H2_M_1224


title: 12H2_M_941


title: 12H2_M_942


title: 12H2_M_959


title: 12H2_M_958


title: 12H2_M_939


title: 12H2_M_953


title: 12H2_M_945


title: 12H2_M_938


title: 12H2_M_946


title: 12H2_M_937


title: 12H2_M_932


title: 12H2_M_933


title: 12H2_M_936


title: 12H2_M_934


title: 12H2_M_929


title: 12H2_M_931


title: 12H2_M_930


title: 12H2_M_940


title: 12H2_M_927


title: 12H2_M_922


title: 12H2_M_925


title: 12H2_M_928


title: 12H2_M_926


title: 12H2_M_923


title: 12H2_M_924


title: 12H2_M_919


title: 12H2_M_921


title: 12H2_M_935


title: 12H2_M_916


title: 12H2_M_915


title: 12H2_M_918


title: 12H2_M_911


title: 12H2_M_914


title: 12H2_M_913


title: 12H2_M_909


title: 12H2_M_917


title: 12H2_M_920


title: 12H2_M_902


title: 12H2_M_906


title: 12H2_M_912


title: 12H2_M_910


title: 12H2_M_907


title: 12H2_M_905


title: 12H2_M_901


title: 12H2_M_903


title: 12H2_M_900


title: 12H2_M_899


title: 12H2_M_894


title: 12H2_M_895


title: 12H2_M_896


title: 12H2_M_897


title: 12H2_M_898


title: 12H2_M_889


title: 12H2_M_892


title: 12H2_M_893


title: 12H2_M_908


title: 12H2_M_890


title: 12H2_M_891


title: 12H2_M_887


title: 12H2_M_884


title: 12H2_M_885


title: 12H2_M_888


title: 12H2_M_883


title: 12H2_M_881


title: 12H2_M_886


title: 12H2_M_904


title: 12H2_M_882


title: 12H2_M_880


title: 12H2_M_874


title: 12H2_M_878


title: 12H2_M_875


title: 12H2_M_879


title: 12H2_M_877


title: 12H2_M_876


title: 12H2_M_861


title: 12H2_M_865


title: 12H2_M_871


title: 12H2_M_868


title: 12H2_M_859


title: 12H2_M_860


title: 12H2_M_864


title: 12H2_M_866


title: 12H2_M_872


title: 12H2_M_855


title: 12H2_M_858


title: 12H2_M_856


title: 12H2_M_870


title: 12H2_M_854


title: 12H2_M_852


title: 12H2_M_857


title: 12H2_M_862


title: 12H2_M_851


title: 12H2_M_853


title: 12H2_M_873


title: 12H2_M_869


title: 12H2_M_867


title: 12H2_M_863


title: 12H2_M_850


title: 12H2_M_844


title: 12H2_M_848


title: 12H2_M_845


title: 12H2_M_847


title: 12H2_M_846


title: 12H2_M_840


title: 12H2_M_839


title: 12H2_M_842


title: 12H2_M_841


title: 12H2_M_843


title: 12H2_M_838


title: 12H2_M_832


title: 12H2_M_835


title: 12H2_M_833


title: 12H2_M_831


title: 12H2_M_823


title: 12H2_M_829


title: 12H2_M_828


title: 12H2_M_830


title: 12H2_M_827


title: 12H2_M_824


title: 12H2_M_834


title: 12H2_M_849


title: 12H2_M_822


title: 12H2_M_825


title: 12H2_M_821


title: 12H2_M_837


title: 12H2_M_820


title: 12H2_M_836


title: 12H2_M_817


title: 12H2_M_816


title: 12H2_M_818


title: 12H2_M_814


title: 12H2_M_810


title: 12H2_M_811


title: 12H2_M_809


title: 12H2_M_812


title: 12H2_M_813


title: 12H2_M_808


title: 12H2_M_804


title: 12H2_M_802


title: 12H2_M_806


title: 12H2_M_805


title: 12H2_M_807


title: 12H2_M_798


title: 12H2_M_797


title: 12H2_M_796


title: 12H2_M_800


title: 12H2_M_819


title: 12H2_M_801


title: 12H2_M_803


title: 12H2_M_799


title: 12H2_M_795


title: 12H2_M_792


title: 12H2_M_794


title: 12H2_M_793


title: 12H2_M_791


title: 12H2_M_815


title: 12H2_M_790


title: 12H2_M_789


title: 12H2_M_788


title: 12H2_M_787


title: 12H2_M_786


title: 12H2_M_782


title: 12H2_M_783


title: 12H2_M_785


title: 12H2_M_781


title: 12H2_M_784


title: 12H2_M_774


title: 12H2_M_777


title: 12H2_M_776


title: 12H2_M_779


title: 12H2_M_778


title: 12H2_M_775


title: 12H2_M_771


title: 12H2_M_773


title: 12H2_M_780


title: 12H2_M_768


title: 12H2_M_575


title: 12H2_M_772


title: 12H2_M_770


title: 12H2_M_573


title: 12H2_M_571


title: 12H2_M_574


title: 12H2_M_769


title: 12H2_M_570


title: 12H2_M_572


title: 12H2_M_568


title: 12H2_M_566


title: 12H2_M_567


title: 12H2_M_569


title: 12H2_M_565


title: 12H2_M_563


title: 12H2_M_564


title: 12H2_M_560


title: 12H2_M_559


title: 12H2_M_561


title: 12H2_M_558


title: 12H2_M_557


title: 12H2_M_556


title: 12H2_M_553


title: 12H2_M_555


title: 12H2_M_543


title: 12H2_M_546


title: 12H2_M_541


title: 12H2_M_544


title: 12H2_M_539


title: 12H2_M_551


title: 12H2_M_552


title: 12H2_M_540


title: 12H2_M_545


title: 12H2_M_548


title: 12H2_M_547


title: 12H2_M_550


title: 12H2_M_542


title: 12H2_M_536


title: 12H2_M_537


title: 12H2_M_535


title: 12H2_M_538


title: 12H2_M_534


title: 12H2_M_549


title: 12H2_M_533


title: 12H2_M_532


title: 12H2_M_554


title: 12H2_M_528


title: 12H2_M_530


title: 12H2_M_529


title: 12H2_M_527


title: 12H2_M_531


title: 12H2_M_526


title: 12H2_M_525


title: 12H2_M_522


title: 12H2_M_524


title: 12H2_M_523


title: 12H2_M_520


title: 12H2_M_519


title: 12H2_M_518


title: 12H2_M_521


title: 12H2_M_517


title: 12H2_M_514


title: 12H2_M_516


title: 12H2_M_447


title: 12H2_M_513


title: 12H2_M_445


title: 12H2_M_515


title: 12H2_M_512


title: 12H2_M_443


title: 12H2_M_442


title: 12H2_M_446


title: 12H2_M_444


title: 12H2_M_440


title: 12H2_M_439


title: 12H2_M_435


title: 12H2_M_438


title: 12H2_M_441


title: 12H2_M_433


title: 12H2_M_437


title: 12H2_M_436


title: 12H2_M_431


title: 12H2_M_429


title: 12H2_M_427


title: 12H2_M_426


title: 12H2_M_432


title: 12H2_M_425


title: 12H2_M_422


title: 12H2_M_423


title: 12H2_M_418


title: 12H2_M_424


title: 12H2_M_420


title: 12H2_M_419


title: 12H2_M_421


title: 12H2_M_428


title: 12H2_M_416


title: 12H2_M_414


title: 12H2_M_415


title: 12H2_M_412


title: 12H2_M_410


title: 12H2_M_411


title: 12H2_M_406


title: 12H2_M_407


title: 12H2_M_405


title: 12H2_M_404


title: 12H2_M_417


title: 12H2_M_398


title: 12H2_M_400


title: 12H2_M_396


title: 12H2_M_401


title: 12H2_M_397


title: 12H2_M_413


title: 12H2_M_388


title: 12H2_M_391


title: 12H2_M_389


title: 12H2_M_392


title: 12H2_M_393


title: 12H2_M_394


title: 12H2_M_390


title: 12H2_M_387


title: 12H2_M_402


title: 12H2_M_409


title: 12H2_M_385


title: 12H2_M_403


title: 12H2_M_384


title: 12H2_M_319


title: 12H2_M_386


title: 12H2_M_395


title: 12H2_M_316


title: 12H2_M_311


title: 12H2_M_313


title: 12H2_M_315


title: 12H2_M_314


title: 12H2_M_309


title: 12H2_M_310


title: 12H2_M_307


title: 12H2_M_308


title: 12H2_M_305


title: 12H2_M_306


title: 12H2_M_303


title: 12H2_M_304


title: 12H2_M_302


title: 12H2_M_301


title: 12H2_M_317


title: 12H2_M_312


title: 12H2_M_297


title: 12H2_M_295


title: 12H2_M_298


title: 12H2_M_296


title: 12H2_M_290


title: 12H2_M_293


title: 12H2_M_294


title: 12H2_M_289


title: 12H2_M_288


title: 12H2_M_285


title: 12H2_M_286


title: 12H2_M_282


title: 12H2_M_283


title: 12H2_M_287


title: 12H2_M_280


title: 12H2_M_279


title: 12H2_M_299


title: 12H2_M_278


title: 12H2_M_430


title: 12H2_M_276


title: 12H2_M_300


title: 12H2_M_275


title: 12H2_M_274


title: 12H2_M_292


title: 12H2_M_291


title: 12H2_M_272


title: 12H2_M_273


title: 12H2_M_271


title: 12H2_M_270


title: 12H2_M_284


title: 12H2_M_281


title: 12H2_M_266


title: 12H2_M_264


title: 12H2_M_263


title: 12H2_M_262


title: 12H2_M_267


title: 12H2_M_268


title: 12H2_M_265


title: 12H2_M_261


title: 12H2_M_260


title: 12H2_M_259


title: 12H2_M_255


title: 12H2_M_256


title: 12H2_M_254


title: 12H2_M_253


title: 12H2_M_257


title: 12H2_M_258


title: 12H2_M_252


title: 12H2_M_277


title: 12H2_M_249


title: 12H2_M_251


title: 12H2_M_250


title: 12H2_M_247


title: 12H2_M_246


title: 12H2_M_245


title: 12H2_M_238


title: 12H2_M_240


title: 12H2_M_241


title: 12H2_M_242


title: 12H2_M_233


title: 12H2_M_243


title: 12H2_M_244


title: 12H2_M_237


title: 12H2_M_234


title: 12H2_M_232


title: 12H2_M_235


title: 12H2_M_236


title: 12H2_M_239


title: 12H2_M_231


title: 12H2_M_248


title: 12H2_M_222


title: 12H2_M_223


title: 12H2_M_225


title: 12H2_M_221


title: 12H2_M_224


title: 12H2_M_226


title: 12H2_M_227


title: 12H2_M_229


title: 12H2_M_217


title: 12H2_M_218


title: 12H2_M_216


title: 12H2_M_215


title: 12H2_M_230


title: 12H2_M_212


title: 12H2_M_210


title: 12H2_M_213


title: 12H2_M_205


title: 12H2_M_204


title: 12H2_M_208


title: 12H2_M_211


title: 12H2_M_214


title: 12H2_M_219


title: 12H2_M_202


title: 12H2_M_203


title: 12H2_M_209


title: 12H2_M_220


title: 12H2_M_200


title: 12H2_M_199


title: 12H2_M_207


title: 12H2_M_193


title: 12H2_M_201


title: 12H2_M_195


title: 12H2_M_194


title: 12H2_M_125


title: 12H2_M_126


title: 12H2_M_192


title: 12H2_M_127


title: 12H2_M_124


title: 12H2_M_206


title: 12H2_M_196


title: 12H2_M_198


title: 12H2_M_118


title: 12H2_M_117


title: 12H2_M_119


title: 12H2_M_197


title: 12H2_M_112


title: 12H2_M_121


title: 12H2_M_120


title: 12H2_M_113


title: 12H2_M_123


title: 12H2_M_122


title: 12H2_M_111


title: 12H2_M_116


title: 12H2_M_114


title: 12H2_M_110


title: 12H2_M_107


title: 12H2_M_109


title: 12H2_M_105


title: 12H2_M_103


title: 12H2_M_106


title: 12H2_M_104


title: 12H2_M_102


title: 12H2_M_100


title: 12H2_M_98


title: 12H2_M_101


title: 12H2_M_99


title: 12H2_M_96


title: 12H2_M_97


title: 12H2_M_115


title: 12H2_M_94


title: 12H2_M_95


title: 12H2_M_93


title: 12H2_M_108


title: 12H2_M_91


title: 12H2_M_92


title: 12H2_M_86


title: 12H2_M_88


title: 12H2_M_85


title: 12H2_M_89


title: 12H2_M_84


title: 12H2_M_82


title: 12H2_M_76


title: 12H2_M_80


title: 12H2_M_81


title: 12H2_M_79


title: 12H2_M_83


title: 12H2_M_78


title: 12H2_M_77


title: 12H2_M_90


title: 12H2_M_75


title: 12H2_M_87


title: 12H2_M_72


title: 12H2_M_74


title: 12H2_M_73


title: 12H2_M_69


title: 12H2_M_71


title: 12H2_M_70


title: 12H2_M_68


title: 12H2_M_67


title: 12H2_M_66


title: 12H2_M_64


title: 12H_M_3066


title: 12H2_M_65


title: 12H_M_3065


title: 12H_M_3064


title: 12H_M_3061


title: 12H_M_3058


title: 12H_M_3060


title: 12H_M_3062


title: 12H_M_3057


title: 12H_M_3063


title: 12H_M_3059


title: 12H_M_3052


title: 12H_M_3051


title: 12H_M_3055


title: 12H_M_3054


title: 12H_M_3056


title: 12H_M_3053


title: 12H_M_3049


title: 12H_M_3048


title: 12H_M_3050


title: 12H_M_3043


title: 12H_M_3044


title: 12H_M_3047


title: 12H_M_3045


title: 12H_M_3046


title: 12H_M_3037


title: 12H_M_3042


title: 12H_M_3038


title: 12H_M_3041


title: 12H_M_3036


title: 12H_M_3040


title: 12H_M_3035


title: 12H_M_3039


title: 12H_M_3032


title: 12H_M_3031


title: 12H_M_3030


title: 12H_M_3033


title: 12H_M_3028


title: 12H_M_3027


title: 12H_M_3024


title: 12H_M_3026


title: 12H_M_3023


title: 12H_M_3021


title: 12H_M_3022


title: 12H_M_3029


title: 12H_M_3020


title: 12H_M_3018


title: 12H_M_3019


title: 12H_M_3034


title: 12H_M_3017


title: 12H_M_3016


title: 12H_M_3008


title: 12H_M_3013


title: 12H_M_3015


title: 12H_M_3011


title: 12H_M_3010


title: 12H_M_3014


title: 12H_M_3012


title: 12H_M_3009


title: 12H_M_3007


title: 12H_M_3006


title: 12H_M_3005


title: 12H_M_3004


title: 12H_M_2873


title: 12H_M_3003


title: 12H_M_2872


title: 12H_M_2874


title: 12H_M_2870


title: 12H_M_2871


title: 12H_M_2869


title: 12H_M_2867


title: 12H_M_2865


title: 12H_M_2866


title: 12H_M_2861


title: 12H_M_2862


title: 12H_M_2863


title: 12H_M_2860


title: 12H_M_2858


title: 12H_M_2859


title: 12H_M_2854


title: 12H_M_2857


title: 12H_M_2853


title: 12H_M_2855


title: 12H_M_2868


title: 12H_M_2851


title: 12H_M_2847


title: 12H_M_2844


title: 12H_M_2850


title: 12H_M_2849


title: 12H_M_2845


title: 12H_M_2856


title: 12H_M_2852


title: 12H_M_2864


title: 12H_M_2842


title: 12H_M_2839


title: 12H_M_2836


title: 12H_M_2846


title: 12H_M_2833


title: 12H_M_2834


title: 12H_M_2843


title: 12H_M_2837


title: 12H_M_2848


title: 12H_M_2831


title: 12H_M_2830


title: 12H_M_2841


title: 12H_M_2826


title: 12H_M_2829


title: 12H_M_2828


title: 12H_M_2827


title: 12H_M_2832


title: 12H_M_2825


title: 12H_M_2822


title: 12H_M_2821


title: 12H_M_2818


title: 12H_M_2819


title: 12H_M_2820


title: 12H_M_2823


title: 12H_M_2816


title: 12H_M_2835


title: 12H_M_2815


title: 12H_M_2813


title: 12H_M_2745


title: 12H_M_2744


title: 12H_M_2741


title: 12H_M_2743


title: 12H_M_2742


title: 12H_M_2740


title: 12H_M_2739


title: 12H_M_2738


title: 12H_M_2817


title: 12H_M_2737


title: 12H_M_2736


title: 12H_M_2734


title: 12H_M_2733


title: 12H_M_2735


title: 12H_M_2732


title: 12H_M_2730


title: 12H_M_2814


title: 12H_M_2812


title: 12H_M_2811


title: 12H_M_2731


title: 12H_M_2727


title: 12H_M_2725


title: 12H_M_2729


title: 12H_M_2746


title: 12H_M_2724


title: 12H_M_2722


title: 12H_M_2721


title: 12H_M_2719


title: 12H_M_2717


title: 12H_M_2718


title: 12H_M_2716


title: 12H_M_2728


title: 12H_M_2726


title: 12H_M_2715


title: 12H_M_2714


title: 12H_M_2712


title: 12H_M_2710


title: 12H_M_2711


title: 12H_M_2713


title: 12H_M_2708


title: 12H_M_2706


title: 12H_M_2704


title: 12H_M_2705


title: 12H_M_2723


title: 12H_M_2703


title: 12H_M_2700


title: 12H_M_2701


title: 12H_M_2699


title: 12H_M_2698


title: 12H_M_2720


title: 12H_M_2697


title: 12H_M_2695


title: 12H_M_2693


title: 12H_M_2694


title: 12H_M_2692


title: 12H_M_2707


title: 12H_M_2709


title: 12H_M_2691


title: 12H_M_2702


title: 12H_M_2690


title: 12H_M_2688


title: 12H_M_2696


title: 12H_M_2684


title: 12H_M_2683


title: 12H_M_2685


title: 12H_M_2687


title: 12H_M_2686


title: 12H_M_2678


title: 12H_M_2676


title: 12H_M_2677


title: 12H_M_2679


title: 12H_M_2680


title: 12H_M_2675


title: 12H_M_2682


title: 12H_M_2671


title: 12H_M_2681


title: 12H_M_2673


title: 12H_M_2674


title: 12H_M_2668


title: 12H_M_2689


title: 12H_M_2669


title: 12H_M_2667


title: 12H_M_2666


title: 12H_M_2665


title: 12H_M_2664


title: 12H_M_2663


title: 12H_M_2662


title: 12H_M_2661


title: 12H_M_2660


title: 12H_M_2670


title: 12H_M_2659


title: 12H_M_2657


title: 12H_M_2655


title: 12H_M_2656


title: 12H_M_2652


title: 12H_M_2653


title: 12H_M_2646


title: 12H_M_2647


title: 12H_M_2650


title: 12H_M_2645


title: 12H_M_2651


title: 12H_M_2640


title: 12H_M_2648


title: 12H_M_2658


title: 12H_M_2641


title: 12H_M_2642


title: 12H_M_2644


title: 12H_M_2639


title: 12H_M_2649


title: 12H_M_2838


title: 12H_M_2637


title: 12H_M_2636


title: 12H_M_2643


title: 12H_M_2635


title: 12H_M_2840


title: 12H_M_2638


title: 12H_M_2634


title: 12H_M_2654


title: 12H_M_2633


title: 12H_M_2631


title: 12H_M_2632


title: 12H_M_2630


title: 12H_M_2629


title: 12H_M_2621


title: 12H_M_2623


title: 12H_M_2622


title: 12H_M_2624


title: 12H_M_2620


title: 12H_M_2625


title: 12H_M_2626


title: 12H_M_2619


title: 12H_M_2614


title: 12H_M_2615


title: 12H_M_2616


title: 12H_M_2617


title: 12H_M_2612


title: 12H_M_2611


title: 12H_M_2609


title: 12H_M_2613


title: 12H_M_2608


title: 12H_M_2610


title: 12H_M_2607


title: 12H_M_2606


title: 12H_M_2605


title: 12H_M_2603


title: 12H_M_2628


title: 12H_M_2627


title: 12H_M_2599


title: 12H_M_2600


title: 12H_M_2618


title: 12H_M_2604


title: 12H_M_2601


title: 12H_M_2598


title: 12H_M_2596


title: 12H_M_2594


title: 12H_M_2592


title: 12H_M_2597


title: 12H_M_2593


title: 12H_M_2589


title: 12H_M_2590


title: 12H_M_2591


title: 12H_M_2586


title: 12H_M_2585


title: 12H_M_2583


title: 12H_M_2588


title: 12H_M_2582


title: 12H_M_2581


title: 12H_M_2580


title: 12H_M_2602


title: 12H_M_2579


title: 12H_M_2576


title: 12H_M_2577


title: 12H_M_2578


title: 12H_M_2575


title: 12H_M_2574


title: 12H_M_2573


title: 12H_M_2595


title: 12H_M_2572


title: 12H_M_2570


title: 12H_M_2571


title: 12H_M_2568


title: 12H_M_2569


title: 12H_M_2567


title: 12H_M_2566


title: 12H_M_2584


title: 12H_M_2587


title: 12H_M_2565


title: 12H_M_2563


title: 12H_M_2562


title: 12H_M_2564


title: 12H_M_2561


title: 12H_M_2557


title: 12H_M_2560


title: 12H_M_2559


title: 12H_M_2558


title: 12H_M_2554


title: 12H_M_2555


title: 12H_M_2556


title: 12H_M_2552


title: 12H_M_2553


title: 12H_M_2549


title: 12H_M_2550


title: 12H_M_2551


title: 12H_M_2548


title: 12H_M_2547


title: 12H_M_2544


title: 12H_M_2545


title: 12H_M_2546


title: 12H_M_2543


title: 12H_M_2541


title: 12H_M_2542


title: 12H_M_2540


title: 12H_M_2538


title: 12H_M_2537


title: 12H_M_2539


title: 12H_M_2535


title: 12H_M_2534


title: 12H_M_2533


title: 12H_M_2536


title: 12H_M_2532


title: 12H_M_2531


title: 12H_M_2530


title: 12H_M_2520


title: 12H_M_2523


title: 12H_M_2522


title: 12H_M_2525


title: 12H_M_2521


title: 12H_M_2527


title: 12H_M_2519


title: 12H_M_2524


title: 12H_M_2528


title: 12H_M_2526


title: 12H_M_2529


title: 12H_M_2516


title: 12H_M_2515


title: 12H_M_2507


title: 12H_M_2508


title: 12H_M_2509


title: 12H_M_2517


title: 12H_M_2510


title: 12H_M_2518


title: 12H_M_2512


title: 12H_M_2511


title: 12H_M_2513


title: 12H_M_2506


title: 12H_M_2505


title: 12H_M_2500


title: 12H_M_2503


title: 12H_M_2504


title: 12H_M_2502


title: 12H_M_2501


title: 12H_M_2514


title: 12H_M_2492


title: 12H_M_2493


title: 12H_M_2498


title: 12H_M_2495


title: 12H_M_2494


title: 12H_M_2427


title: 12H_M_2497


title: 12H_M_2496


title: 12H_M_2499


title: 12H_M_2416


title: 12H_M_2417


title: 12H_M_2415


title: 12H_M_2420


title: 12H_M_2424


title: 12H_M_2414


title: 12H_M_2421


title: 12H_M_2418


title: 12H_M_2413


title: 12H_M_2423


title: 12H_M_2410


title: 12H_M_2408


title: 12H_M_2411


title: 12H_M_2409


title: 12H_M_2407


title: 12H_M_2406


title: 12H_M_2425


title: 12H_M_2426


title: 12H_M_2419


title: 12H_M_2422


title: 12H_M_2403


title: 12H_M_2405


title: 12H_M_2401


title: 12H_M_2404


title: 12H_M_2397


title: 12H_M_2398


title: 12H_M_2402


title: 12H_M_2399


title: 12H_M_2400


title: 12H_M_2412


title: 12H_M_2394


title: 12H_M_2392


title: 12H_M_2396


title: 12H_M_2395


title: 12H_M_2386


title: 12H_M_2387


title: 12H_M_2388


title: 12H_M_2390


title: 12H_M_2391


title: 12H_M_2389


title: 12H_M_2384


title: 12H_M_2385


title: 12H_M_2381


title: 12H_M_2382


title: 12H_M_2383


title: 12H_M_2380


title: 12H_M_2379


title: 12H_M_2378


title: 12H_M_2377


title: 12H_M_2376


title: 12H_M_2371


title: 12H_M_2372


title: 12H_M_2373


title: 12H_M_2375


title: 12H_M_2374


title: 12H_M_2369


title: 12H_M_2393


title: 12H_M_2367


title: 12H_M_2366


title: 12H_M_2368


title: 12H_M_2365


title: 12H_M_2364


title: 12H_M_2362


title: 12H_M_2363


title: 12H_M_2361


title: 12H_M_2359


title: 12H_M_2357


title: 12H_M_2358


title: 12H_M_2356


title: 12H_M_2354


title: 12H_M_2351


title: 12H_M_2352


title: 12H_M_2348


title: 12H_M_2349


title: 12H_M_2350


title: 12H_M_2370


title: 12H_M_2347


title: 12H_M_2346


title: 12H_M_2345


title: 12H_M_2344


title: 12H_M_2360


title: 12H_M_2340


title: 12H_M_2338


title: 12H_M_2339


title: 12H_M_2341


title: 12H_M_2336


title: 12H_M_2353


title: 12H_M_2355


title: 12H_M_2331


title: 12H_M_2334


title: 12H_M_2332


title: 12H_M_2333


title: 12H_M_2329


title: 12H_M_2326


title: 12H_M_2343


title: 12H_M_2324


title: 12H_M_2327


title: 12H_M_2342


title: 12H_M_2328


title: 12H_M_2322


title: 12H_M_2323


title: 12H_M_2337


title: 12H_M_2335


title: 12H_M_2316


title: 12H_M_2321


title: 12H_M_2317


title: 12H_M_2315


title: 12H_M_2330


title: 12H_M_2313


title: 12H_M_2314


title: 12H_M_2312


title: 12H_M_2310


title: 12H_M_2311


title: 12H_M_2308


title: 12H_M_2309


title: 12H_M_2306


title: 12H_M_2307


title: 12H_M_2305


title: 12H_M_2325


title: 12H_M_2304


title: 12H_M_2301


title: 12H_M_2303


title: 12H_M_2320


title: 12H_M_2318


title: 12H_M_2300


title: 12H_M_2319


title: 12H_M_2295


title: 12H_M_2294


title: 12H_M_2293


title: 12H_M_2299


title: 12H_M_2298


title: 12H_M_2292


title: 12H_M_2290


title: 12H_M_2291


title: 12H_M_2288


title: 12H_M_2289


title: 12H_M_2287


title: 12H_M_2283


title: 12H_M_2286


title: 12H_M_2285


title: 12H_M_2284


title: 12H_M_2282


title: 12H_M_2277


title: 12H_M_2280


title: 12H_M_2281


title: 12H_M_2279


title: 12H_M_2276


title: 12H_M_2278


title: 12H_M_2297


title: 12H_M_2271


title: 12H_M_2269


title: 12H_M_2274


title: 12H_M_2272


title: 12H_M_2273


title: 12H_M_2270


title: 12H_M_2268


title: 12H_M_2266


title: 12H_M_2265


title: 12H_M_2264


title: 12H_M_2267


title: 12H_M_2263


title: 12H_M_2262


title: 12H_M_2261


title: 12H_M_2258


title: 12H_M_2260


title: 12H_M_2275


title: 12H_M_2257


title: 12H_M_2259


title: 12H_M_2256


title: 12H_M_2254


title: 12H_M_2253


title: 12H_M_2250


title: 12H_M_2249


title: 12H_M_2252


title: 12H_M_2251


title: 12H_M_2247


title: 12H_M_2248


title: 12H_M_2246


title: 12H_M_2245


title: 12H_M_2243


title: 12H_M_2241


title: 12H_M_2244


title: 12H_M_2240


title: 12H_M_2242


title: 12H_M_2237


title: 12H_M_2238


title: 12H_M_2170


title: 12H_M_2171


title: 12H_M_2239


title: 12H_M_2172


title: 12H_M_2169


title: 12H_M_2167


title: 12H_M_2168


title: 12H_M_2165


title: 12H_M_2166


title: 12H_M_2255


title: 12H_M_2162


title: 12H_M_2161


title: 12H_M_2159


title: 12H_M_2164


title: 12H_M_2158


title: 12H_M_2156


title: 12H_M_2155


title: 12H_M_2154


title: 12H_M_2153


title: 12H_M_2152


title: 12H_M_2151


title: 12H_M_2150


title: 12H_M_2148


title: 12H_M_2149


title: 12H_M_2144


title: 12H_M_2143


title: 12H_M_2145


title: 12H_M_2141


title: 12H_M_2142


title: 12H_M_2140


title: 12H_M_2147


title: 12H_M_2163


title: 12H_M_2146


title: 12H_M_2136


title: 12H_M_2138


title: 12H_M_2160


title: 12H_M_2157


title: 12H_M_2137


title: 12H_M_2135


title: 12H_M_2134


title: 12H_M_2131


title: 12H_M_2139


title: 12H_M_2129


title: 12H_M_2127


title: 12H_M_2126


title: 12H_M_2125


title: 12H_M_2124


title: 12H_M_2118


title: 12H_M_2122


title: 12H_M_2121


title: 12H_M_2117


title: 12H_M_2119


title: 12H_M_2133


title: 12H_M_2132


title: 12H_M_2120


title: 12H_M_2115


title: 12H_M_2111


title: 12H_M_2112


title: 12H_M_2123


title: 12H_M_2130


title: 12H_M_1979


title: 12H_M_2110


title: 12H_M_2116


title: 12H_M_2114


title: 12H_M_1980


title: 12H_M_2128


title: 12H_M_1977


title: 12H_M_2113


title: 12H_M_1978


title: 12H_M_1975


title: 12H_M_1976


title: 12H_M_2109


title: 12H_M_1971


title: 12H_M_1973


title: 12H_M_1974


title: 12H_M_1970


title: 12H_M_1972


title: 12H_M_1968


title: 12H_M_1969


title: 12H_M_1967


title: 12H_M_1965


title: 12H_M_1964


title: 12H_M_1966


title: 12H_M_1963


title: 12H_M_1960


title: 12H_M_1961


title: 12H_M_1962


title: 12H_M_1954


title: 12H_M_1957


title: 12H_M_1953


title: 12H_M_1956


title: 12H_M_1958


title: 12H_M_1955


title: 12H_M_1950


title: 12H_M_1951


title: 12H_M_1952


title: 12H_M_1959


title: 12H_M_1949


title: 12H_M_1947


title: 12H_M_1948


title: 12H_M_1942


title: 12H_M_1943


title: 12H_M_1944


title: 12H_M_1945


title: 12H_M_1946


title: 12H_M_1940


title: 12H_M_1941


title: 12H_M_1939


title: 12H_M_1937


title: 12H_M_1938


title: 12H_M_1934


title: 12H_M_1936


title: 12H_M_1935


title: 12H_M_1933


title: 12H_M_1932


title: 12H_M_1931


title: 12H_M_1927


title: 12H_M_1930


title: 12H_M_1929


title: 12H_M_1928


title: 12H_M_1926


title: 12H_M_1925


title: 12H_M_1921


title: 12H_M_1923


title: 12H_M_1924


title: 12H_M_1919


title: 12H_M_1922


title: 12H_M_1918


title: 12H_M_1914


title: 12H_M_1912


title: 12H_M_1916


title: 12H_M_1915


title: 12H_M_1913


title: 12H_M_1917


title: 12H_M_1911


title: 12H_M_1907


title: 12H_M_1908


title: 12H_M_1910


title: 12H_M_1906


title: 12H_M_1904


title: 12H_M_1920


title: 12H_M_1902


title: 12H_M_1901


title: 12H_M_1903


title: 12H_M_1900


title: 12H_M_1897


title: 12H_M_1899


title: 12H_M_1892


title: 12H_M_1891


title: 12H_M_1894


title: 12H_M_1896


title: 12H_M_1889


title: 12H_M_1888


title: 12H_M_1886


title: 12H_M_1883


title: 12H_M_1885


title: 12H_M_1905


title: 12H_M_1890


title: 12H_M_1884


title: 12H_M_1909


title: 12H_M_1882


title: 12H_M_1887


title: 12H_M_1881


title: 12H_M_1879


title: 12H_M_1878


title: 12H_M_1876


title: 12H_M_1880


title: 12H_M_1898


title: 12H_M_1895


title: 12H_M_1877


title: 12H_M_1893


title: 12H_M_1873


title: 12H_M_1874


title: 12H_M_1872


title: 12H_M_1870


title: 12H_M_1869


title: 12H_M_1871


title: 12H_M_1865


title: 12H_M_1868


title: 12H_M_1864


title: 12H_M_1867


title: 12H_M_1863


title: 12H_M_1862


title: 12H_M_1866


title: 12H_M_1858


title: 12H_M_1857


title: 12H_M_1861


title: 12H_M_1875


title: 12H_M_1860


title: 12H_M_1859


title: 12H_M_1854


title: 12H_M_1856


title: 12H_M_1851


title: 12H_M_1849


title: 12H_M_1850


title: 12H_M_1852


title: 12H_M_1855


title: 12H_M_1853


title: 12H_M_1848


title: 12H_M_1847


title: 12H_M_1846


title: 12H_M_1840


title: 12H_M_1838


title: 12H_M_1845


title: 12H_M_1843


title: 12H_M_1834


title: 12H_M_1836


title: 12H_M_1837


title: 12H_M_1839


title: 12H_M_1833


title: 12H_M_1831


title: 12H_M_1832


title: 12H_M_1830


title: 12H_M_1825


title: 12H_M_1828


title: 12H_M_1842


title: 12H_M_1827


title: 12H_M_1822


title: 12H_M_1823


title: 12H_M_1824


title: 12H_M_1829


title: 12H_M_1844


title: 12H_M_1841


title: 12H_M_1817


title: 12H_M_1818


title: 12H_M_1816


title: 12H_M_1821


title: 12H_M_1815


title: 12H_M_1807


title: 12H_M_1810


title: 12H_M_1814


title: 12H_M_1826


title: 12H_M_1820


title: 12H_M_1812


title: 12H_M_1803


title: 12H_M_1806


title: 12H_M_1805


title: 12H_M_1797


title: 12H_M_1813


title: 12H_M_1802


title: 12H_M_1796


title: 12H_M_1798


title: 12H_M_1819


title: 12H_M_1795


title: 12H_M_1794


title: 12H_M_1792


title: 12H_M_1811


title: 12H_M_1793


title: 12H_M_1791


title: 12H_M_1800


title: 12H_M_1808


title: 12H_M_1804


title: 12H_M_1809


title: 12H_M_1801


title: 12H_M_1789


title: 12H_M_1790


title: 12H_M_1799


title: 12H_M_1724


title: 12H_M_1721


title: 12H_M_1723


title: 12H_M_1720


title: 12H_M_1722


title: 12H_M_1715


title: 12H_M_1714


title: 12H_M_1717


title: 12H_M_1716


title: 12H_M_1709


title: 12H_M_1712


title: 12H_M_1710


title: 12H_M_1713


title: 12H_M_1707


title: 12H_M_1711


title: 12H_M_1708


title: 12H_M_1704


title: 12H_M_1705


title: 12H_M_1706


title: 12H_M_1702


title: 12H_M_1701


title: 12H_M_1700


title: 12H_M_1703


title: 12H_M_1718


title: 12H_M_1719


title: 12H_M_1697


title: 12H_M_1699


title: 12H_M_1698


title: 12H_M_1696


title: 12H_M_1694


title: 12H_M_1693


title: 12H_M_1691


title: 12H_M_1690


title: 12H_M_1692


title: 12H_M_1688


title: 12H_M_1686


title: 12H_M_1687


title: 12H_M_1689


title: 12H_M_1682


title: 12H_M_1683


title: 12H_M_1684


title: 12H_M_1685


title: 12H_M_1679


title: 12H_M_1695


title: 12H_M_1680


title: 12H_M_1681


title: 12H_M_1675


title: 12H_M_1678


title: 12H_M_1677


title: 12H_M_1676


title: 12H_M_1672


title: 12H_M_1669


title: 12H_M_1674


title: 12H_M_1673


title: 12H_M_1671


title: 12H_M_1670


title: 12H_M_1668


title: 12H_M_1666


title: 12H_M_1667


title: 12H_M_1665


title: 12H_M_1662


title: 12H_M_1663


title: 12H_M_1664


title: 12H_M_1405


title: 12H_M_1406


title: 12H_M_1661


title: 12H_M_1404


title: 12H_M_1403


title: 12H_M_1401


title: 12H_M_1402


title: 12H_M_1400


title: 12H_M_1399


title: 12H_M_1398


title: 12H_M_1397


title: 12H_M_1394


title: 12H_M_1396


title: 12H_M_1392


title: 12H_M_1390


title: 12H_M_1385


title: 12H_M_1383


title: 12H_M_1395


title: 12H_M_1387


title: 12H_M_1388


title: 12H_M_1389


title: 12H_M_1384


title: 12H_M_1381


title: 12H_M_1380


title: 12H_M_1376


title: 12H_M_1391


title: 12H_M_1371


title: 12H_M_1372


title: 12H_M_1374


title: 12H_M_1373


title: 12H_M_1375


title: 12H_M_1386


title: 12H_M_1393


title: 12H_M_1370


title: 12H_M_1369


title: 12H_M_1368


title: 12H_M_1382


title: 12H_M_1365


title: 12H_M_1364


title: 12H_M_1379


title: 12H_M_1359


title: 12H_M_1366


title: 12H_M_1361


title: 12H_M_1360


title: 12H_M_1378


title: 12H_M_1358


title: 12H_M_1357


title: 12H_M_1362


title: 12H_M_1356


title: 12H_M_1355


title: 12H_M_1354


title: 12H_M_1353


title: 12H_M_1349


title: 12H_M_1350


title: 12H_M_1351


title: 12H_M_1347


title: 12H_M_1343


title: 12H_M_1346


title: 12H_M_1345


title: 12H_M_1367


title: 12H_M_1363


title: 12H_M_1081


title: 12H_M_1086


title: 12H_M_1083


title: 12H_M_1084


title: 12H_M_1080


title: 12H_M_1085


title: 12H_M_1078


title: 12H_M_1079


title: 12H_M_1075


title: 12H_M_1077


title: 12H_M_1074


title: 12H_M_1072


title: 12H_M_1076


title: 12H_M_1073


title: 12H_M_1348


title: 12H_M_1070


title: 12H_M_1071


title: 12H_M_1068


title: 12H_M_1067


title: 12H_M_1069


title: 12H_M_1065


title: 12H_M_1064


title: 12H_M_1082


title: 12H_M_1063


title: 12H_M_1066


title: 12H_M_1062


title: 12H_M_1352


title: 12H_M_1061


title: 12H_M_1059


title: 12H_M_1057


title: 12H_M_1060


title: 12H_M_1056


title: 12H_M_1054


title: 12H_M_1058


title: 12H_M_1049


title: 12H_M_1051


title: 12H_M_1053


title: 12H_M_1055


title: 12H_M_1052


title: 12H_M_1047


title: 12H_M_1050


title: 12H_M_1046


title: 12H_M_1045


title: 12H_M_1044


title: 12H_M_1043


title: 12H_M_1042


title: 12H_M_1041


title: 12H_M_1039


title: 12H_M_1040


title: 12H_M_1036


title: 12H_M_1035


title: 12H_M_1037


title: 12H_M_1038


title: 12H_M_1034


title: 12H_M_1031


title: 12H_M_1048


title: 12H_M_1344


title: 12H_M_1033


title: 12H_M_1032


title: 12H_M_1030


title: 12H_M_1029


title: 12H_M_1835


title: 12H_M_1024


title: 12H_M_1025


title: 12H_M_1026


title: 12H_M_1027


title: 12H_M_1028


title: 12H_M_1023


title: 12H_M_1022


title: 12H_M_1015


title: 12H_M_1019


title: 12H_M_1010


title: 12H_M_1009


title: 12H_M_1012


title: 12H_M_1020


title: 12H_M_1014


title: 12H_M_1011


title: 12H_M_1017


title: 12H_M_1004


title: 12H_M_1005


title: 12H_M_1006


title: 12H_M_1007


title: 12H_M_1003


title: 12H_M_1021


title: 12H_M_1002


title: 12H_M_1001


title: 12H_M_1018


title: 12H_M_998


title: 12H_M_1013


title: 12H_M_999


title: 12H_M_997


title: 12H_M_1016


title: 12H_M_1000


title: 12H_M_995


title: 12H_M_996


title: 12H_M_994


title: 12H_M_1008


title: 12H_M_992


title: 12H_M_1377


title: 12H_M_988


title: 12H_M_993


title: 12H_M_990


title: 12H_M_987


title: 12H_M_989


title: 12H_M_991


title: 12H_M_984


title: 12H_M_982


title: 12H_M_986


title: 12H_M_985


title: 12H_M_981


title: 12H_M_983


title: 12H_M_980


title: 12H_M_978


title: 12H_M_979


title: 12H_M_977


title: 12H_M_976


title: 12H_M_975


title: 12H_M_973


title: 12H_M_972


title: 12H_M_971


title: 12H_M_970


title: 12H_M_974


title: 12H_M_967


title: 12H_M_969


title: 12H_M_968


title: 12H_M_966


title: 12H_M_965


title: 12H_M_964


title: 12H_M_962


title: 12H_M_961


title: 12H_M_963


title: 12H_M_959


title: 12H_M_958


title: 12H_M_956


title: 12H_M_947


title: 12H_M_957


title: 12H_M_951


title: 12H_M_950


title: 12H_M_954


title: 12H_M_948


title: 12H_M_952


title: 12H_M_955


title: 12H_M_949


title: 12H_M_946


title: 12H_M_944


title: 12H_M_940


title: 12H_M_943


title: 12H_M_945


title: 12H_M_938


title: 12H_M_941


title: 12H_M_936


title: 12H_M_953


title: 12H_M_960


title: 12H_M_935


title: 12H_M_937


title: 12H_M_934


title: 12H_M_933


title: 12H_M_931


title: 12H_M_929


title: 12H_M_932


title: 12H_M_924


title: 12H_M_925


title: 12H_M_942


title: 12H_M_926


title: 12H_M_920


title: 12H_M_921


title: 12H_M_918


title: 12H_M_939


title: 12H_M_919


title: 12H_M_917


title: 12H_M_915


title: 12H_M_912


title: 12H_M_923


title: 12H_M_910


title: 12H_M_913


title: 12H_M_914


title: 12H_M_930


title: 12H_M_908


title: 12H_M_909


title: 12H_M_928


title: 12H_M_905


title: 12H_M_927


title: 12H_M_904


title: 12H_M_901


title: 12H_M_900


title: 12H_M_902


title: 12H_M_922


title: 12H_M_896


title: 12H_M_899


title: 12H_M_897


title: 12H_M_898


title: 12H_M_916


title: 12H_M_911


title: 12H_M_895


title: 12H_M_894


title: 12H_M_890


title: 12H_M_891


title: 12H_M_892


title: 12H_M_907


title: 12H_M_889


title: 12H_M_888


title: 12H_M_887


title: 12H_M_884


title: 12H_M_885


title: 12H_M_886


title: 12H_M_880


title: 12H_M_883


title: 12H_M_881


title: 12H_M_879


title: 12H_M_882


title: 12H_M_877


title: 12H_M_878


title: 12H_M_876


title: 12H_M_874


title: 12H_M_875


title: 12H_M_873


title: 12H_M_871


title: 12H_M_893


title: 12H_M_868


title: 12H_M_869


title: 12H_M_866


title: 12H_M_861


title: 12H_M_863


title: 12H_M_862


title: 12H_M_870


title: 12H_M_867


title: 12H_M_860


title: 12H_M_857


title: 12H_M_864


title: 12H_M_859


title: 12H_M_865


title: 12H_M_855


title: 12H_M_858


title: 12H_M_856


title: 12H_M_853


title: 12H_M_854


title: 12H_M_852


title: 12H_M_850


title: 12H_M_851


title: 12H_M_872


title: 12H_M_849


title: 12H_M_848


title: 12H_M_845


title: 12H_M_846


title: 12H_M_843


title: 12H_M_844


title: 12H_M_842


title: 12H_M_839


title: 12H_M_840


title: 12H_M_841


title: 12H_M_838


title: 12H_M_837


title: 12H_M_835


title: 12H_M_836


title: 12H_M_834


title: 12H_M_832


title: 12H_M_638


title: 12H_M_833


title: 12H_M_831


title: 12H_M_636


title: 12H_M_637


title: 12H_M_635


title: 12H_M_632


title: 12H_M_633


title: 12H_M_630


title: 12H_M_627


title: 12H_M_634


title: 12H_M_629


title: 12H_M_631


title: 12H_M_625


title: 12H_M_626


title: 12H_M_621


title: 12H_M_622


title: 12H_M_618


title: 12H_M_617


title: 12H_M_623


title: 12H_M_615


title: 12H_M_628


title: 12H_M_609


title: 12H_M_604


title: 12H_M_608


title: 12H_M_614


title: 12H_M_610


title: 12H_M_612


title: 12H_M_613


title: 12H_M_605


title: 12H_M_606


title: 12H_M_603


title: 12H_M_602


title: 12H_M_607


title: 12H_M_601


title: 12H_M_599


title: 12H_M_598


title: 12H_M_597


title: 12H_M_624


title: 12H_M_619


title: 12H_M_600


title: 12H_M_616


title: 12H_M_620


title: 12H_M_611


title: 12H_M_594


title: 12H_M_592


title: 12H_M_593


title: 12H_M_595


title: 12H_M_590


title: 12H_M_589


title: 12H_M_596


title: 12H_M_584


title: 12H_M_583


title: 12H_M_582


title: 12H_M_585


title: 12H_M_588


title: 12H_M_587


title: 12H_M_586


title: 12H_M_581


title: 12H_M_579


title: 12H_M_577


title: 12H_M_578


title: 12H_M_580


title: 12H_M_591


title: 12H_M_445


title: 12H_M_443


title: 12H_M_442


title: 12H_M_575


title: 12H_M_441


title: 12H_M_444


title: 12H_M_437


title: 12H_M_438


title: 12H_M_446


title: 12H_M_576


title: 12H_M_439


title: 12H_M_440


title: 12H_M_436


title: 12H_M_433


title: 12H_M_434


title: 12H_M_432


title: 12H_M_430


title: 12H_M_421


title: 12H_M_418


title: 12H_M_431


title: 12H_M_416


title: 12H_M_429


title: 12H_M_422


title: 12H_M_417


title: 12H_M_425


title: 12H_M_428


title: 12H_M_424


title: 12H_M_423


title: 12H_M_408


title: 12H_M_414


title: 12H_M_427


title: 12H_M_407


title: 12H_M_409


title: 12H_M_410


title: 12H_M_412


title: 12H_M_419


title: 12H_M_420


title: 12H_M_405


title: 12H_M_403


title: 12H_M_435


title: 12H_M_413


title: 12H_M_411


title: 12H_M_402


title: 12H_M_406


title: 12H_M_404


title: 12H_M_415


title: 12H_M_401


title: 12H_M_399


title: 12H_M_397


title: 12H_M_396


title: 12H_M_398


title: 12H_M_394


title: 12H_M_392


title: 12H_M_393


title: 12H_M_391


title: 12H_M_390


title: 12H_M_385


title: 12H_M_381


title: 12H_M_382


title: 12H_M_383


title: 12H_M_389


title: 12H_M_384


title: 12H_M_377


title: 12H_M_387


title: 12H_M_375


title: 12H_M_388


title: 12H_M_379


title: 12H_M_378


title: 12H_M_380


title: 12H_M_376


title: 12H_M_373


title: 12H_M_374


title: 12H_M_372


title: 12H_M_400


title: 12H_M_395


title: 12H_M_371


title: 12H_M_370


title: 12H_M_369


title: 12H_M_368


title: 12H_M_364


title: 12H_M_361


title: 12H_M_365


title: 12H_M_363


title: 12H_M_360


title: 12H_M_358


title: 12H_M_357


title: 12H_M_355


title: 12H_M_353


title: 12H_M_352


title: 12H_M_354


title: 12H_M_350


title: 12H_M_356


title: 12H_M_348


title: 12H_M_349


title: 12H_M_346


title: 12H_M_367


title: 12H_M_343


title: 12H_M_344


title: 12H_M_345


title: 12H_M_342


title: 12H_M_341


title: 12H_M_347


title: 12H_M_340


title: 12H_M_366


title: 12H_M_362


title: 12H_M_359


title: 12H_M_338


title: 12H_M_339


title: 12H_M_337


title: 12H_M_334


title: 12H_M_336


title: 12H_M_335


title: 12H_M_351


title: 12H_M_333


title: 12H_M_330


title: 12H_M_327


title: 12H_M_329


title: 12H_M_332


title: 12H_M_328


title: 12H_M_325


title: 12H_M_323


title: 12H_M_331


title: 12H_M_322


title: 12H_M_324


title: 12H_M_321


title: 12H_M_326


title: 12H_M_319


title: 12H_M_316


title: 12H_M_315


title: 12H_M_317


title: 12H_M_318


title: 12H_M_314


title: 12H_M_320


title: 12H_M_313


title: 12H_M_311


title: 12H_M_308


title: 12H_M_309


title: 12H_M_310


title: 12H_M_307


title: 12H_M_305


title: 12H_M_312


title: 12H_M_304


title: 12H_M_302


title: 12H_M_301


title: 12H_M_300


title: 12H_M_303


title: 12H_M_299


title: 12H_M_298


title: 12H_M_296


title: 12H_M_297


title: 12H_M_295


title: 12H_M_292


title: 12H_M_291


title: 12H_M_294


title: 12H_M_290


title: 12H_M_289


title: 12H_M_284


title: 12H_M_283


title: 12H_M_287


title: 12H_M_306


title: 12H_M_288


title: 12H_M_286


title: 12H_M_282


title: 12H_M_279


title: 12H_M_276


title: 12H_M_275


title: 12H_M_273


title: 12H_M_293


title: 12H_M_272


title: 12H_M_271


title: 12H_M_270


title: 12H_M_269


title: 12H_M_262


title: 12H_M_265


title: 12H_M_261


title: 12H_M_267


title: 12H_M_257


title: 12H_M_258


title: 12H_M_285


title: 12H_M_260


title: 12H_M_263


title: 12H_M_277


title: 12H_M_281


title: 12H_M_255


title: 12H_M_253


title: 12H_M_256


title: 12H_M_252


title: 12H_M_278


title: 12H_M_274


title: 12H_M_268


title: 12H_M_259


title: 12H_M_254


title: 12H_M_251


title: 12H_M_250


title: 12H_M_247


title: 12H_M_249


title: 12H_M_266


title: 12H_M_246


title: 12H_M_241


title: 12H_M_243


title: 12H_M_242


title: 12H_M_264


title: 12H_M_236


title: 12H_M_240


title: 12H_M_235


title: 12H_M_238


title: 12H_M_234


title: 12H_M_237


title: 12H_M_239


title: 12H_M_233


title: 12H_M_280


title: 12H_M_248


title: 12H_M_231


title: 12H_M_228


title: 12H_M_245


title: 12H_M_229


title: 12H_M_232


title: 12H_M_230


title: 12H_M_225


title: 12H_M_223


title: 12H_M_227


title: 12H_M_226


title: 12H_M_222


title: 12H_M_220


title: 12H_M_224


title: 12H_M_221


title: 12H_M_218


title: 12H_M_219


title: 12H_M_216


title: 12H_M_210


title: 12H_M_217


title: 12H_M_215


title: 12H_M_213


title: 12H_M_206


title: 12H_M_209


title: 12H_M_208


title: 12H_M_207


title: 12H_M_203


title: 12H_M_212


title: 12H_M_205


title: 12H_M_204


title: 12H_M_198


title: 12H_M_197


title: 12H_M_202


title: 12H_M_200


title: 12H_M_199


title: 12H_M_201


title: 12H_M_192


title: 12H_M_194


title: 12H_M_195


title: 12H_M_193


title: 12H_M_196


title: 12H_M_191


title: 12H2_M_1343


title: 12H2_M_1342


title: 12H_M_211


title: 12H2_M_1341


title: 12H2_M_1336


title: 12H2_M_1333


title: 12H2_M_1338


title: 12H2_M_1334


title: 12H2_M_1337


title: 12H2_M_1340


title: 12H2_M_1335


title: 12H2_M_1331


title: 12H2_M_1328


title: 12H2_M_1329


title: 12H2_M_1332


title: 12H2_M_1326


title: 12H2_M_1325


title: 12H2_M_1324


title: 12H2_M_1327


title: 12H2_M_1322


title: 12H2_M_1321


title: 12H2_M_1320


title: 12H2_M_1319


title: 12H2_M_1315


title: 12H2_M_1317


title: 12H2_M_1316


title: 12H2_M_1318


title: 12H2_M_1314


title: 12H2_M_1313


title: 12H2_M_1339


title: 12H2_M_1309


title: 12H2_M_1308


title: 12H2_M_1305


title: 12H2_M_1304


title: 12H2_M_1306


title: 12H2_M_1303


title: 12H2_M_1311


title: 12H2_M_1300


title: 12H2_M_1302


title: 12H2_M_1323


title: 12H2_M_1301


title: 12H2_M_1298


title: 12H2_M_1299


title: 12H2_M_1295


title: 12H2_M_1294


title: 12H2_M_1291


title: 12H2_M_1293


title: 12H2_M_1292


title: 12H2_M_1312


title: 12H2_M_1288


title: 12H2_M_1290


title: 12H2_M_1286


title: 12H2_M_1310


title: 12H2_M_1287


title: 12H2_M_1285


title: 12H2_M_1307


title: 12H2_M_1284


title: 12H2_M_1283


title: 12H2_M_1282


title: 12H2_M_1215


title: 12H2_M_1297


title: 12H2_M_1281


title: 12H2_M_1280


title: 12H2_M_1213


title: 12H2_M_1214


title: 12H2_M_1210


title: 12H2_M_1208


title: 12H2_M_1296


title: 12H2_M_1207


title: 12H2_M_1212


title: 12H2_M_1211


title: 12H2_M_1206


title: 12H2_M_1209


title: 12H2_M_1205


title: 12H2_M_1204


title: 12H2_M_1201


title: 12H2_M_1200


title: 12H2_M_1202


title: 12H2_M_1196


title: 12H2_M_1197


title: 12H2_M_1195


title: 12H2_M_1194


title: 12H2_M_1198


title: 12H2_M_1199


title: 12H2_M_1193


title: 12H2_M_1192


title: 12H2_M_1191


title: 12H2_M_1190


title: 12H2_M_1188


title: 12H2_M_1184


title: 12H2_M_1185


title: 12H2_M_1187


title: 12H2_M_1189


title: 12H2_M_1182


title: 12H2_M_1183


title: 12H2_M_1179


title: 12H2_M_1203


title: 12H2_M_1181


title: 12H2_M_1176


title: 12H2_M_1178


title: 12H2_M_1177


title: 12H2_M_1175


title: 12H2_M_1173


title: 12H2_M_1174


title: 12H2_M_1168


title: 12H2_M_1171


title: 12H2_M_1169


title: 12H2_M_1170


title: 12H2_M_1167


title: 12H2_M_1172