title: 4sU-seq_HepG2_JTE-607_2

data_processing:
(1). MPIVA: All MPIVA read 1 and read 2 FASTQ files were merged using bbmerge V38 using option ` maxloose=t`.
(1). MPIVA: MPIVA Input samples were used establish the sequences of the full randomized PAS region contained in the IVT pool by clustering the 25 nt randomized region with starcode version 1.4. The expected cleaved lengths of the 25 nt consensus sequences for L3 and SVL backbones (13 nt and 12 nt, respectively) were used as unique identifiers of the full randomized region. If any of these identifiers were not unique within L3 and SVL libraries, respectively, then these sequences were not used in subsequent analyses.
(1). MPIVA: RNA-seq reads from DMSO and drug-treated libraries were locally aligned against the shared 5′ region of the reporter constructs to determine the beginning of the randomized region. The part of the RNA-seq read containing the randomized region and shared 3′ region was locally aligned against the list of consensus sequences from the Input libraries.
(1). MPIVA: Sequences with at least 50 reads in the DMSO libraries were kept to avoid noise from lowly abundant RNA sequences in the IVT pool.
(1). MPIVA: A pseudocount of 1 was added to variants in L3 2.5 µM, L3 12.5 µM, and SVL 12.5 µM due to drug-mediated drop out of high abundance variants in the DMSO libraries. This pseudocount avoids having undefined drug sensitivities in later steps that would be introduced by log(0).
(1). MPIVA: Each variant that passed these filters were counted and converted to a percentage within each RNA-seq library.
(1). MPIVA: Drug sensitivity for each variant in each dose of Compound 2 was defined as the log ratio of normalized reads from drug-treated RNA-seq divided by the normalized reads from DMSO-treated RNA-seq.
(2). 4sU-seq: Reads were mapped to human(hg19) genome using STAR. Multi-mapping was not allowed. Mapped reads were converted to bigwig files using deepTools.
(3). PAS-seq: Reads that lack a poly(A) tail (15 consecutive As) were removed with python script. The trimmed reads were mapped to human (hg19) genome using STAR.
(3). PAS-seq: Removed internal priming reads (reads that are mapped to genomic sequences that are immediately followed by 6-consecutive As, or seven As out of 10 nucleotides)
(3). PAS-seq: Reads were converted to BigWig files using deepTools
Assembly: hg19
Supplementary files format and content: (1). MPIVA: TSV file containing log ratio of different concentrations of Compound 2 vs DMSO in both L3 and SVL backbone and their sequences.
Supplementary files format and content: (2). 4sU-seq and (3). PAS-seq: BigWig files.
Library strategy: 4sU-seq

value: 4sU labeled total RNA


title: Nephrotoxic Nephritis sample of wild type mouse (replicate 2)

status: Public on Jul 22 2023

type: SRA

city: Cologne

first: Ali

middle: T

last: Abdallah

address: Joseph Stelzmann Str. 26

department: Medical Faculty

country: Germany

institute: University of Cologne

library_source: transcriptomic single cell

data_processing:
Libraries were sequenced on an Illumina NovaSeq 6000 sequencing instrument with 29+89bp read length.
Fastqs were generated using the command mkfastq of the 10x Genomics software "cellranger" (v6.1.2). The filtered gene count matrices of the four samples were generated using cellranger's count command and the mm10 reference genome (version preprocessed by cellranger). For the two WT and the two BtlaKO samples, 3.5K, 5.4K, 6.4K, and 8.4K cells were recovered, respectively. Sequencing depths of the samples were about 64.9K, 60.3K, 52.2K, and 37.2K reads per cell in the same order. That resulted in a median number of genes per cell between 2K and 2.3K for each sample. All given numbers were rounded to the first decimal place.
Downstream analysis was performed using the Seurat R package (version 4.1.1) 29. Cells with a very small (<500 genes) or large (>5000 genes) transcriptome recovery were discarded to reduce noise and the number of possible doublets, respectively. Only cells with mitochondrial expression of less than 5% and ribosomal expression of less than 50% were kept to avoid working with low-quality cells in downstream steps. For normalization, we have used the standard Log-normalization approach with the standard scaling factor (10e4). Samples were then integrated using the standard Seurat integration procedure.
We followed the standard Seurat-workflow to perform principal component analysis, extracting the first 100 principal components. Based on the data generated by Seurat's Elbow plot function, we automatically determined the number of dimensions used for dimensionality reduction and clustering of all immune cells. In particular, we selected the dimension where the standard deviation starts to drop below 2; this automatic selection procedure was confirmed by visual inspection of the elbow plot. Subsequently, we have executed the UMAP algorithm and the FindNeighbors function with the aforementioned dimensions. Clustering was performed using the FindClusters function with a resolution of 0.1. Otherwise, we used standard parameters. The dimensionality reduction and clustering procedure for the T cells was performed similarly, but we used only the first 12 dimensions by visual inspection of the elbow plot, as the complexity and variability within T cell population drops as expected significantly compared to first level of resolution (all immune cells). In this step we used a clustering resolution of 0.6. Cluster 6 was identified as contamination with macrophages, probably due to the formation of doublets and was discarded. The other cells were reclustered using the same workflow, this time with a clustering resolution of 0.5. For all downstream steps we worked with the remaining 5412, 7361, 2939, and 4521 cells for the knockout and wild-type samples, respectively. At T cell level, we worked accordingly with 826, 1433, 366, and 609 cells in the same mentioned order.
Markers of the clusters at both clustering levels were computed with the Wilcoxon rank sum test implemented in Seurat using the FindAllMarkers function. The top 20 positive markers of each cluster were used to manually curate the cell types of the immune cells (the initial clustering level of all cells). The reference database used to determine the cell types is the single-cell RNASeq database PanglaoDB30. We determined the identity of the resulting cluster subpopulations by visualizing the expression patterns of most basic markers (e.g., Cd4+, Cd8+, and some alpha, beta, gamma, and delta chain genes) across clusters to understand the approximate structure of T cell subpopulations in our dataset. Subsequently, we have computed the module scores of many known standard subpopulations of T cells for each cell in each cluster using the AddModuleScore function. By box plotting the distribution of the scores of each module for each cluster, we identify the subpopulations of the T cells by visually inspecting and comparing the module score distributions. To assign a cell type to a subpopulation we require that the median score is at least higher than zero or that no other type is fitting better while the median being close to zero. In the case that two cell type modules are fitting equally well into the same subcluster, we annotate the cluster with both unless we could biologically exclude one of them. For instance, in the case of clusters where almost all cells are Cd4+, we exclude the possibility of the cluster being a subpopulation of cytotoxic T cells. Cycling T cells were recognized using cell cycle genes integrated within Seurat and corresponding G2M and S scores. The markers of the various T cell subpopulations were taken from the Biocompare website
Assembly: mm10 (preprocessed release downloaded from the website of 10x Genomics)
Supplementary files format and content: four csv files for the raw counts generated by cellranger for each sample and three additional csv files; the first for the integrated and filtered object from the four raw samples and the other two contain the cell type annotation of all immune cells and the fine-granuled annotation of the T-cells, respectively

molecule: total RNA

organism: Mus musculus

source_name: Wild type

growth_protocol: Not applicable

extract_protocol:
kidneys where digested, CD45+ leukocytes isolated using MACS beads, then sorted using FACS gating on Living (DAPIneg),CD45+
10xGenomics 3`scRNAseq

treatment_protocol: Induction of nephrotoxic nephritis 10 days prior to organ harvest

tag: cell type

value: renal leukocytes

tag: treatment

value: Wild type

tag: tissue

value: Kidney

tag: disease state

value: Nephrotoxic Nephritis


title: FKBP5high - rep2

city: Amsterdam

first: Isabel

last: Mayayo-Peralta

address: Plesmanlaan 121

country: Netherlands

institute: NKI

library_source: genomic

data_processing:
Read counts were normalized, relative total sizefactors were calculated and that the values within a sample were divided by the respective sample sizefactor
reads of each replicate were pooled together and subsequent analyses were performed using MaGeCK, using default parameters (v0.5.9.4)
Putative GR regulators were identified comparing the sgRNA abundance amongst the 7.5% FKBP5high vs 7.5% FKBP5low populations and a robust rank aggregation (RRA) score was determined for each gene
Those genes with an FDR<0.15, log2(fold change)>2.5 and –log10(p-value)>2 were considered as putative hits.
Assembly: GRCh38.102
Supplementary files format and content: readcounts, normalized readcounts, mageck results
Library strategy: DNA-seq / CRISPR screen

molecule: genomic DNA

organism: Homo sapiens

growth_protocol: DMEM/F12 (1:1) (1x) + Glutamax + 10% FCS + 1% Pen/Strep, cultured at 5% CO2 and 37 °C.

extract_protocol: After treatment, cells were fixed in 70% ethanol and left overnight at 4 °C. Fixed cells were washed with PBS and permeabilized with PBS containing 0.25% triton X-100 (Sigma-Aldrich) and left on ice for 15 min. Cells were stained with a recombinant anti-FKBP5 anti-body (AlexaFluor 647, ab198979, 1:400) in 1% bovine serum albumin (BSA)-PBS. Stained cells were washed with PBS and resuspended in 1%BSA-PBS. Cells were then FACS sorted for 7.5% FKBP5high or 7.5% FKBP5low of the population, with ~3.500.000 cells/arm to ensure a 500x coverage of the library. Genomic DNA was subsequently isolated using Gentra Pura-gene Cell kit according to manufacturer’s instructions. gRNAs were amplified by two consecutive PCRs as previously described (Korkmaz et al., 2016).

treatment_protocol: The Brunello CRISPR lentiviral library was transduced into A549 cells at a low multiplicity of transduction (MOI) of ~0.3 to ensure that only one sgRNA was incorporated per cell. Transduced cells were subsequently selected with 2μg/mL of puromycin for 3 days and left untreated for 14 days. After that, cells were incubated in DMEM/F12 (1:1) (1X) + Glutamax, supplemented with dextran-coated charcoal-treated FCS (DCC) for 24 hours and then treat-ed with 1μg/mL of hydrocortisone (HC) for additional 24 hours

value: Lung Cancer

tag: cell line

tag: genotype

value: WT

value: 1ug/mL hydrocortisone

tag: treatment length

tag: facs antibody

value: FKBP5 antibody (AlexaFluor 647, ab198979)


title: Nephrotoxic Nephritis sample of wild type mouse (replicate 1)


title: 4sU-seq_HepG2_JTE-607_1

city: Irvine

first: Yongsheng

last: Shi

address: 811 Health Sciences Rd, Med Sci B262

country: USA

institute: University of California, Irvine

library_source: transcriptomic

source_name: liver

extract_protocol:
(1). MPIVA: RNA from in vitro assay is phenol chloroform extracted and polyadenylated RNA is purified using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB).
(2). 4sU-seq and (3). PAS-seq: RNA is extracted by Trizol. For 4sU-seq, RNA is biotinylated and 4sU labeled RNA is enriched by strepavdin beads and further purified by phenol chloroform extraction
(1). MPIVA: Library is constructed by RT-PCR.
(2). 4sU-seq and (3). PAS-seq: RNA is fragmented and library is constructed following illumina's protocol

treatment_protocol:
(1). MPIVA: Libraries were cloned by gibson assembly and in vitro transcribed to RNA. The RNA pool was then used in in vitro cleavage and polyadenylation assay.
(2). 4sU-seq: HepG2 cells were treated with DMSO or 20µM JTE-607 for 3 hours at 37˚C followed by addition of 500µM 4sU. The cell containing JTE-607 and 4sU was incubated at 37˚C for another 1 hour prior to sample harvesting.
(3). PAS-seq: HepG2 cells were treated with DMSO or 20µM JTE-607 for 4 hours at 37˚C prior to sample harvesting

value: liver

value: human liver carcinoma cell

tag: sample type


title: Nephrotoxic Nephritis sample of BTLA-KO mouse (replicate 2)

source_name: Knock out

value: Knock-out


title: FKBP5high - rep1


title: Mock-1

city: Melbourne

first: Kevin

middle: C

last: sek

state: VIC

address: 305 Grattan Street

department: Cancer Immunology

country: Australia

institute: Peter MacCallum Cancer Centre

data_processing:
Illumina Casava1.8.2 software used for basecalling.
Reads were trimmed for adaptor sequence using Ngmerge, then mapped to the mm10 genome with bowtie2
Peaks were called with Macs2
Assembly: mm10
Supplementary files format and content: Peak files called with macs2

extract_protocol:
Gene edited AT-3 breast cancer cells lysed using the ZYMO research DNA isolation kit. Tagmentation then purification was performed on DNA, and isolated chromatin was amplified.
Sequencing libraries were constructed using the Illumina Nextera DNA Sample prep kit.

value: Breast Cancer

value: Mock edited

value: stimulation with IFNgamma and TNF


title: 4sU-seq_HepG2_DMSO_2

value: DMSO


title: Nephrotoxic Nephritis sample of BTLA-KO mouse (replicate 1)


title: FKBP5low - rep3


title: SOCS1-KO-2


title: 4sU-seq_HepG2_DMSO_1


title: FKBP5low - rep1


title: C8

city: Guangzhou

first: Yifang

last: Gao

address: NO.58 zhongshan 2nd road, YueXiu district

country: China

institute: sun yat-sen University

description:
Healthy liver
matrix.mtx
barcodes.tsv
genes.tsv

data_processing:
Basecalls performed using CASAVA version 1.8.
Cellranger was utilized to calculate the raw counts and cell whitelist by default parameter.
We applied the Seurat [https://satijalab.org/seurat/] package for cell normalization and cell filtering considering the MT percentage, minimum and maximum gene numbers under following criteria:
o MT% < 20%;
o Cell Gene Number < Median Gene Number * 2;
o Cell Gene Number > 200.
PCA and tSNE analysis was used for the single cell to cell relation description. The fastMNN function (k = 5, d = 50, approximate = TRUE, auto.order = TRUE) from R package scran (v1.10.2) was used to apply mutual nearest neighbor method to correct for batch effect among samples. Graphcluster was utilized for cell clustering and based on the marker gene achieved some cluster was combined. Wilcox rank sum test was then used for marker gene analysis.
Genome_build: GRCh38_Ensembl91
Supplementary_files_format_and_content: MTX file and TSV file contain counts for each barcodes and genes.

source_name: liver perfusate

growth_protocol: Cells were isolated from perfusates of liver that were going to transplant.

extract_protocol:
Single-cell gel beads in emulsion (GEMs) were generated on a GemCode Single Cell Instrument (10x Genomics). Single-cell RNA-seq cDNA libraries were prepared using Chromium™ Single Cell 3' Reagent Kit v3 (10x Genomics) according to manufacturer's protocol.
RNA libraries were generated by 10x Genomics Single Cell 3’ Library Gel Bead Kit V2 and prepared for sequencing using standard Illumina protocols.

treatment_protocol: Cells were extracted by density gradient centrifugation with Ficoll-Paque Plus.

tag: diagnosis

value: Healthy

tag: tissue type

value: Liver


title: HOTTIP-KO-AML-CTCF-ChIP-seq-MOLM13

city: Hangzhou

first: Huacheng

last: Luo

state: Zhejiang

address: No. 150, Fucheng Road, Qiantang District

institute: Hangzhou Institute of Medicine (HIM), The Chinese Academy of Sciences

description: HOTTIP-KO-AML-CTCF-ChIP-seq

data_processing:
Standard Illumina software base-calling and quality-control filtering was applied Sequences (Paired end)
Paired end HiC-seq and scRNA-seq and scATAC-seq reads from mice were aligned to the mm9 genome assembly using BWA, and NG capture-C-seq, ChIP-seq, DRIP-seq and DRIPc-seq reads from human cells were aligned to the hg19 genome assembly using BOWTIE2 default parameters.
Peak calling was performed using MACS algorithm (version 2.1.1)
Genome coverage tracks were created using deepTools version 3.0
Assembly: hg19
Supplementary files format and content: tab-delimited text files include FPKM values for scRNA-seq and scATAC-seq Samples and peaks for DRIP-seq , DRIPc-seq, ChIP-seq, NG-capture-C and HIC-seq.

source_name: peripheral blood

growth_protocol: Total bone marrow cells were isolate from WT and Hottip-Tg mice, and sorted with Lin-, Sca1+ and Kit+ markers.  Human MOLM13 leukemia cells were culture at RMPI1640 medium with 10% FBS media.

extract_protocol:
RNA samples extracted with Trizon and treated with Dnase I.  ChIP Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with CTCF, SA1, SA2, RAD21, H3K4me3 and H3K27me3 antibody (antibody: Anti-CTCF, Cell Signaling Technology, Cat# 2899; Anti-SA1, Abcam, Cat# ab4457; Anti-SA2, Abcam, Cat# ab4464;  Anti-RAD21, Abcam, Cat# ab992;  Anti-H3K4me3, Millipore, Cat No. 04-745;   Anti-H3K27me3, Millipore, Cat No. 07-449).  ATAC-seq samples derived according to Nextera Tn5 Transposase kits. DNA:RNA immunoprecipitation sequencing (DRIP-seq) was performed as described previously with s9.6 antibody (Anti-DNA-RNA Hybrid [S9.6] Antibody, Kerafast, Cat# ENH001).  NG-Capture-C assay was performed as previously described. Hi-C assay was performed to generate a genome-wide interaction as described previously with Arima-HiC Kit (Cat: A410030) (https://arimagenomics.com/) with minor modifications. Single cells were generated using Chromium Controller (10x Genomics), and scRNA-seq and scATAC-seq libraries were constructed using chromium single cell 3’ reagent kits v2 (10x Genomics, California USA) according to the manufacturer’s recommendations.
RNA-seq libraries were prepared according to TruSeq Stranded mRNA Library Prep (#20020594). ChIP-seq libraries were prepared according to Illumina's instructions accompanying the TruSeq ChIP Library Preparation Kit (#IP-202-1012). ATAC-seq libraries were prepared according to Nextera DNA Library Prep Kit (#FC-121-1030). NG-Capture-C-seq libraries were prepared according to using the Herculase II PCR kit (Agilent). . Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.

treatment_protocol: Immediately extract RNA from LK, LSK cells and MOLM13 AML cells without any treatment

value: MOLM13 leukemia cells

value: Human-derived acute myeloid leukemia

value: peripheral blood

value: acute myeloid leukemia

tag: chip antibody

value: Cell Signaling Technology, Cat# 2899

value: HOTTIP KO


title: WT-AML-CTCF-ChIP-seq

description: WT-AML-CTCF-ChIP-seq

value: WT


title: PiKP-785 cells, Kras* 3d off

city: Houston

first: Kalluri

last: Lab

state: Texas

address: 1901 East Rd

institute: UT MD Anderson Cancer Center

description: 10x Genomics

data_processing:
The demultiplexing, barcoded processing, gene counting and aggregation were made using the Cell Ranger software v2.1.0 (https://support.10xgenomics.com/single-cell-gene-expression/software/pipelines/latest/what-is-cell-ranger)
Assembly: mm10
Supplementary files format and content: Tab-separated values files and matrix files

source_name: Pancreatic ductal adenocarcinoma

extract_protocol:
The PiKP-785 primary cell line was cultured in RPMI-1640, 10% Tet System Approved FBS (Tet-free FBS), and 1x penicillin-streptomycin (PS). Cells were cultured in medium with 1 μg/mL Dox to ensure oncogenic Kras expression (Kras* on). To extinguish oncogenic Kras, Dox was withdrawn from the media (Kras* off). Cells were washed in PBS, trypsinized (0.25% trypsin in PBS) and submitted in 10% tet-free FBS RPMI to the Advanced Technology Genomics Core, MDACC.
Single cell Gel Bead-In-Emulsions generation and barcoding, post GEM-RT cleanup and cDNA amplification, library construction and Illumina-ready sequencing library generation were prepared by following the manufacturer’s guidelines. cDNA concentration was estimated using High Sensitivity dsDNA Qubit kit and quantified using HS DNA Bioanalyzer. Following capture and lysis, cDNA was synthesized and amplified to construct Illumina sequencing libraries and sequenced with Illumina Nextseq 500.

value: Pancreatic ductal adenocarcinoma

value: Ductal

value: PiKP

value: Dox 3d off


title: PiKP-785 cells, Kras* 5d off

description: 10x Genomics

value: Dox 5d off


title: D8

description:
HCC tumor
matrix.mtx
barcodes.tsv
genes.tsv

value: HCC

value: Tumor


title: TC20, 9 root

status: Public on Jul 21 2023

city: Tucson

first: Priyanka

last: Kushwaha

state: Arizona

department: Environmental Science

institute: University of Arizona

data_processing:
Read quality was checked using FastQC.
Sequence reads were trimmed for adaptor sequence/low-quality sequence using TrimGalore.
Trimmed reads were checked and erroneous K-mers were removed using Rcorrector.
De-novo assembly of transcriptome was carried out using Trinity.
Kallisto was used to align and quanitfy transcripts in each sample.
Transcripts were mapped against BLAST database against Arabidopsis thaliana genome.
Differential gene expression analysis was performed on genes that were mapped against A. thaliana using DESeq2.
Functional annotation of genes was performed using UniProt and ePlant.
Assembly: GCF_000001735.4
Supplementary files format and content: tab-delimited text files include normalized counts generated using Deseq2

organism: Atriplex lentiformis

source_name: root

extract_protocol:
RNA was harvested using Rneasy Plant mini kit (Qiagen). 500 ng of total RNA was used for the construction of sequencing libraries.
RNA libraries for RNA-seq were prepared using TruSeq Stranded Total RNA sample preparation kit following manufacturer's protocols.

tag: plant

value: Atriplex lentiformis (Torr.) S. Wats.

value: root

value: 20% compost + mine tailings

tag: time

value: 11 weeks

tag: plant height_cm

tag: number of_branches

tag: length longest_branch_cm

tag: plant stress

tag: zinc root_conc_ug_g

tag: arsenic root_conc_ug_g

tag: lead root_conc_ug_g


title: PiKP-785 cells, Kras* On

description: 10x Genomics

tag: cell line

tag: cell type

tag: treatment

value: Dox On


title: TC15, 10 root

tag: tissue

value: 15% compost + mine tailings


title: TC20, 5 root


title: TC15, 6 root


title: TC10, 4 root

value: 10% compost + mine tailings


title: TC15, 2 root


title: TC20, 3 root


title: Inf5

city: České Budějovice

first: Adam

last: Bajgar

state: CZ

address: Branisovska 1645/31a

department: Molecular biology and Genetics

country: Czech Republic

institute: University of South Bohemia

data_processing:
PairEnd transcriptomic data were tested for  content of exons, length of transcripts, read quality and mapping, (CLC Genomics Wokbench 21.0.5)
Data were mapped by using DragenRNA pipeline, Illumina,
Adapter triming - "Trim Reads" tool
Trimmed reads were aligned to reference D. melanogaster genome BDGP6.95 (Ensembl release), annotation is based on FlyBase data base.
Based on PCA analysis, samples Con2_S2_L001 and INf2_S7_L001 were excluded from subsequent analyses
Assembly: D. melanogaster genome BDGP6.95 (Ensembl release)
Supplementary files format and content: tab-delimited text files

molecule: polyA RNA

organism: Drosophila melanogaster

source_name: Plasmatocytes

extract_protocol:
FACS (details provided in associated publication, RNA was isolated by using TriZol according to standard protocol, quality of RNA was analyzed by Bioanalyzer and alll samples passed test with RIN higher than 7
mRNA fraction of total RNA was obtained by depletion of ribozomal RNAs by using siTOOLs riboPOOL D. melanogaster RNA kit (EastPort) followed by NEBNext Ultra II Directional RNA kit (Illumina) fomr preparation of libraries

treatment_protocol: Adult flies were injected by either 5nl of PBS or 50nl of PBS containing 20 thousand of streptococci, imune cells fronm these flies were isolated by FACS by using Crq>GFP  signal.

value: Plasmatocytes

tag: genotype

value: Crq-Gal4; UAS-GFP

value: Inf


title: Inf3


title: TC10, 2 root


title: Inf4


title: Inf2


title: PiKP-785 cells, Kras* On off on

description: 10x Genomics

value: Pancreatic ductal adenocarcinoma

value: Dox On off on


title: TC10, 5 root


title: Inf1


title: FKBP5high - rep3

value: Lung Cancer

value: WT

value: 1ug/mL hydrocortisone

tag: treatment length

tag: facs antibody

value: FKBP5 antibody (AlexaFluor 647, ab198979)


title: Con5

value: Con


title: FKBP5low - rep2


title: Con2


title: Con7


title: Con3


title: Skin_HC_3

city: St. Louis

first: Aaron

last: Ver Heul

state: MO

address: 660 S Euclid Ave.

department: Medicine

institute: Washington University School of Medicine

description: gene expression

data_processing:
Basecalls and demultiplexing were performed with Illumina’s bcl2fastq2 software
RNA-sequencing reads were aligned to the Ensembl release 76 assembly with STAR v2.0.4b (Dobin et al., 2013).
Gene counts were derived from the number of uniquely aligned unambiguous reads by Subread:featureCount v1.4.5 (Liao et al., 2014).
Transcript counts were produced by Sailfish v0.6.3 (Patro et al., 2014).
Sequencing performance was assessed for total number of aligned reads, total number of uniquely aligned reads, genes and transcripts detected, ribosomal fraction, known junction saturation, and read distribution over known gene models with RSeQC v2.3 (Wang et al., 2012).
Assembly: Homo_sapiens reference build Ensembl_R76
Supplementary files format and content: Text files containing transcript counts quantified using Sailfish

source_name: skin

extract_protocol:
Human skin tissues were obtained as 4 mm punch biopsies or de-identified surgical tissue and stored in RNAlater at -80C before processing. To extract whole tissue RNA, samples were homogenized using a bead homogenizer and processed using the QIAGEN RNeasy Kit following the manufacturer’s instructions. Following total RNA extraction, samples were treated with DNase (Turbo DNA-Free Kit, Invitrogen) following the manufacturer’s instructions.
Library preparation was performed with 1 ug of total RNA, and RNA integrity was determined using an Agilent Bioanalyzer. Ribosomal RNA was removed by a hybridization method using Ribo-ZERO kits (Illumina). mRNA was then fragmented in buffer containing 40 mM Tris acetate (pH 8.2), 100 mM potassium acetate, and 30 mM magnesium acetate and heated to 94C for 150 s. mRNA was reverse transcribed to yield cDNA using SuperScript III RT enzyme (Invitrogen, per manufacturer’s instructions) and random hexamers. A second strand reaction was performed to yield ds-cDNA. cDNA was blunt ended, had an A base added to the 3 prime ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 12 cycles using primers incorporating unique index tags.

value: skin

tag: condition

value: HC


title: Skin_AD_4

description: gene expression

value: AD


title: Con1


title: Skin_CPUO_4

description: gene expression

value: CPUO


title: Skin_HC_4

description: gene expression


title: Skin_HC_2

description: gene expression


title: Skin_CPUO_3

description: gene expression


title: Skin_AD_3

description: gene expression


title: Skin_CPUO_2

description: gene expression


title: Skin_HC_1

description: gene expression


title: Skin_CPUO_1

description: gene expression


title: 14_HEK_Diff_92hs

city: Bethesda

first: Ramiro

last: Iglesias-Bartolome

address: 37 Convent Drive

department: LCMB

institute: NIH/NCI

data_processing:
Basecalling, RTA 1.18.66.3, Illumina instrument run time analysis software
Demultiplexing, Bcl2fastq 2.17, Barcode demultiplexed allowing 1 mismatch
Filtering (Adaptor and quality), Trimmomatic 0.36, PE -threads 16 -phred33 ILLUMINACLIP:TruSeq_and_nextera_adapters.fa:2:36:10 LEADING:10 TRAILING:10 MAXINFO:50:0.97 MINLEN:20
Alignment, STAR 2.5.1, --genomeDir $star_genome --readFilesIn R1_trimmed.fastq R2_trimmed.fastq --outTmpDir /logs/star --outSAMunmapped Within --outFilterType BySJout --outFilterMultimapNmax 20 --outFilterMismatchNmax 999 --outFilterMismatchNoverLmax 0.04 --alignIntronMin 20 --alignIntronMax 1000000 --alignMatesGapMax 1000000 --alignSJoverhangMin 8 --alignSJDBoverhangMin 1 --sjdbScore 1 --runThreadN 8 --genomeLoad NoSharedMemory --outSAMtype BAM Unsorted --quantMode TranscriptomeSAM
Gene counts were calculated using Quantify to annotation model with Partek Flow v7
counts were filtered to exclude genes with maximum <= 5.0 with Partek Flow v7
Counts were added +1 and gene expression was normalized to TMM and TPM using PartekFlow v7
TMM values were used for differential analysis to identify genes that are differentially expressed using the GSA function in PartekFlow v7 with q value calculation, by comparing each group to HEK_Basal.
Assembly: hg19
Supplementary files format and content: TMM_all_samples.xlsx
Supplementary files format and content: TMM_TPM_GSEA_all_samples.xlsx

growth_protocol: Cells were cultured at 37°C in the presence of 5% CO2 and cultured in Lonza KGM Keratinocyte Growth Medium BulletKit

extract_protocol:
RNA was isolated and processed using RNeasy Plus Mini Kit (Qiagen) according to manufacturer’s instructions. Cells were lysed using the Precellys lysing kit (Bertin Instruments).
TruSeq mRNA Prep Kit RS-122-2101

treatment_protocol:
For differentiation conditions (Diff), cells were plated under confluent conditions. 48hs after plating media was supplemented with 1.5mM calcium and cells were harvested 24, 48 or 92 hs later. Control non-differentiated cells (Basal) were plated in non confluent conditions and harvested 48hs after plating.
For no epinephrine (Basal-NoEpi) conditions, cells were cultured for two passages (approximately 6 days), without the addition of Epinephrine to basal culture media. Plated in non confluent conditions and harvested 48hs after plating.

value: Primary neonatal pooled human keratinocytes

tag: replicates


title: 15_HEK_Diff_92hs


title: 11_HEK_Diff_48hs


title: 8_HEK_Diff_24hs


title: 6_HEK_Basal_NoEpi

value: No Epinephrine


title: 9_HEK_Diff_24hs


title: 7_HEK_Diff_24hs


title: 5_HEK_Basal_NoEpi


title: 10_HEK_Diff_48hs


title: 12_HEK_Diff_48hs


title: 4_HEK_Basal_NoEpi


title: 3_HEK_Basal


title: 2_HEK_Basal


title: 1_HEK_Basal


title: Skin_AD_2

description: gene expression


title: 13_HEK_Diff_92hs


title: 9L2_M_1532

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-seq2

source_name: C57BL/6J Male P56 Mouse Brain

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

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

tag: brain region

tag: age

tag: strain


title: 9L2_M_1534


title: 9L2_M_1530


title: 9L2_M_1521


title: 9L2_M_1533


title: Skin_AD_1

description: gene expression


title: 9L2_M_1520


title: 9L2_M_1529


title: 9L2_M_1522


title: 9L2_M_1523


title: 9L2_M_1524


title: 9L2_M_1535


title: 9L2_M_1528


title: 9L2_M_1526


title: 9L2_M_1515


title: 9L2_M_1516


title: 9L2_M_1517


title: 9L2_M_1531


title: 9L2_M_1518


title: 9L2_M_1508


title: 9L2_M_1513


title: 9L2_M_1512


title: 9L2_M_1510


title: 9L2_M_1511


title: 9L2_M_1525


title: 9L2_M_1509


title: 9L2_M_1505


title: 9L2_M_1514


title: 9L2_M_1507


title: 9L2_M_1499


title: 9L2_M_1519


title: 9L2_M_1496


title: 9L2_M_1504


title: 9L2_M_1501


title: 9L2_M_1492


title: 9L2_M_1495


title: 9L2_M_1498


title: 9L2_M_1482


title: 9L2_M_1486


title: 9L2_M_1491


title: 9L2_M_1487


title: 9L2_M_1480


title: 9L2_M_1488


title: 9L2_M_1481


title: 9L2_M_1478


title: 9L2_M_1484


title: 9L2_M_1476


title: 9L2_M_1475


title: 9L2_M_1474


title: 9L2_M_1473


title: 9L2_M_1470


title: 9L2_M_1469


title: 9L2_M_1497


title: 9L2_M_1471


title: 9L2_M_1494


title: 9L2_M_1468


title: 9L2_M_1467


title: 9L2_M_1485


title: 9L2_M_1493


title: 9L2_M_1477


title: 9L2_M_1464


title: 9L2_M_1465


title: 9L2_M_1463


title: 9L2_M_1462


title: 9L2_M_1466


title: 9L2_M_1458


title: 9L2_M_1460


title: 9L2_M_1459


title: 9L2_M_1455


title: 9L2_M_1452


title: 9L2_M_1454


title: 9L2_M_1457


title: 9L2_M_1456


title: 9L2_M_1448


title: 9L2_M_1453


title: 9L2_M_1449


title: 9L2_M_1450


title: 9L2_M_1447


title: 9L2_M_1446


title: 9L2_M_1444


title: 9L2_M_1443


title: 9L2_M_1442


title: 9L2_M_1441


title: 9L2_M_1445


title: 9L2_M_1438


title: 9L2_M_1490


title: 9L2_M_1436


title: 9L2_M_1437


title: 9L2_M_1439


title: 9L2_M_1430


title: 9L2_M_1433


title: 9L2_M_1434


title: 9L2_M_1432


title: 9L2_M_1435


title: 9L2_M_1431


title: 9L2_M_1451


title: 9L2_M_1440


title: 9L2_M_1429


title: 9L2_M_1425


title: 9L2_M_1426


title: 9L2_M_1428


title: 9L2_M_1420


title: 9L2_M_1427


title: 9L2_M_1421


title: 9L2_M_1422


title: 9L2_M_1418


title: 9L2_M_1419


title: 9L2_M_1416


title: 9L2_M_1417


title: 9L2_M_1415


title: 9L2_M_1413


title: 9L2_M_1424


title: 9L2_M_1412


title: 9L2_M_1423


title: 9L2_M_1408


title: 9L2_M_1407


title: 9L2_M_1409


title: 9L2_M_1403


title: 9L2_M_1402


title: 9L2_M_1406


title: 9L2_M_1400


title: 9L2_M_1404


title: 9L2_M_1399


title: 9L2_M_1397


title: 9L2_M_1395


title: 9L2_M_1396


title: 9L2_M_1401


title: 9L2_M_1391


title: 9L2_M_1389


title: 9L2_M_1394


title: 9L2_M_1388


title: 9L2_M_1414


title: 9L2_M_1398


title: 9L2_M_1393


title: 9L2_M_1387


title: 9L2_M_1390


title: 9L2_M_1386


title: 9L2_M_1385


title: 9L2_M_1384


title: 9L2_M_1405


title: 9L2_M_1410


title: 9L2_M_1383


title: 9L2_M_1382


title: 9L2_M_1381


title: 9L2_M_1379


title: 9L2_M_1378


title: 9L2_M_1380


title: 9L2_M_1392


title: 9L2_M_1411


title: 9L2_M_1374


title: 9L2_M_1375


title: 9L2_M_1376


title: 9L2_M_1370


title: 9L2_M_1368


title: 9L2_M_1367


title: 9L2_M_1371


title: 9L2_M_1365


title: 9L2_M_1362


title: 9L2_M_1360


title: 9L2_M_1361


title: 9L2_M_1366


title: 9L2_M_1364


title: 9L2_M_1359


title: 9L2_M_1372


title: 9L2_M_1357


title: 9L2_M_1377


title: 9L2_M_1356


title: 9L2_M_1355


title: 9L2_M_1373


title: 9L2_M_1354


title: 9L2_M_1363


title: 9L2_M_1369


title: 9L2_M_1346


title: 9L2_M_1344


title: 9L2_M_1349


title: 9L2_M_1350


title: 9L2_M_1348


title: 9L2_M_1351


title: 9L2_M_1343


title: 9L2_M_1358


title: 9L2_M_1342


title: 9L2_M_1338


title: 9L2_M_1353


title: 9L2_M_1339


title: 9L2_M_1341


title: 9L2_M_1352


title: 9L2_M_1337


title: 9L2_M_1336


title: 9L2_M_1335


title: 9L2_M_1333


title: 9L2_M_1328


title: 9L2_M_1330


title: 9L2_M_1325


title: 9L2_M_1326


title: 9L2_M_1327


title: 9L2_M_1323


title: 9L2_M_1324


title: 9L2_M_1322


title: 9L2_M_1321


title: 9L2_M_1319


title: 9L2_M_1316


title: 9L2_M_1318


title: 9L2_M_1314


title: 9L2_M_1340


title: 9L2_M_1313


title: 9L2_M_1317


title: 9L2_M_1310


title: 9L2_M_1312


title: 9L2_M_1311


title: 9L2_M_1309


title: 9L2_M_1308


title: 9L2_M_1332


title: 9L2_M_1307


title: 9L2_M_1305


title: 9L2_M_1331


title: 9L2_M_1306


title: 9L2_M_1304


title: 9L2_M_1302


title: 9L2_M_1303


title: 9L2_M_1296


title: 9L2_M_1294


title: 9L2_M_1297


title: 9L2_M_1300


title: 9L2_M_1301


title: 9L2_M_1295


title: 9L2_M_1288


title: 9L2_M_1298


title: 9L2_M_1292


title: 9L2_M_1287


title: 9L2_M_1291


title: 9L2_M_1293


title: 9L2_M_1286


title: 9L2_M_1281


title: 9L2_M_1290


title: 9L2_M_1320


title: 9L2_M_1285


title: 9L2_M_1282


title: 9L2_M_1283


title: 9L2_M_1284


title: 9L2_M_1276


title: 9L2_M_1274


title: 9L2_M_1279


title: 9L2_M_1278


title: 9L2_M_1275


title: 9L2_M_1273


title: 9L2_M_1271


title: 9L2_M_1277


title: 9L2_M_1272


title: 9L2_M_1270


title: 9L2_M_1267


title: 9L2_M_1264


title: 9L2_M_1269


title: 9L2_M_1268


title: 9L2_M_1299


title: 9L2_M_1261


title: 9L2_M_1263


title: 9L2_M_1280


title: 9L2_M_1266


title: 9L2_M_1256


title: 9L2_M_1262


title: 9L2_M_1260


title: 9L2_M_1259


title: 9L2_M_1252


title: 9L2_M_1258


title: 9L2_M_1251


title: 9L2_M_1253


title: 9L2_M_1255


title: 9L2_M_1254


title: 9L2_M_1250


title: 9L2_M_1257


title: 9L2_M_1249


title: 9L2_M_1247


title: 9L2_M_1248


title: 9L2_M_1265


title: 9L2_M_1246


title: 9L2_M_1244


title: 9L2_M_1245


title: 9L2_M_1238


title: 9L2_M_1242


title: 9L2_M_1241


title: 9L2_M_1239


title: 9L2_M_1240


title: 9L2_M_1233


title: 9L2_M_1237


title: 9L2_M_1230


title: 9L2_M_1235


title: 9L2_M_1231


title: 9L2_M_1229


title: 9L2_M_1232


title: 9L2_M_1226


title: 9L2_M_1225


title: 9L2_M_1223


title: 9L2_M_1228


title: 9L2_M_1221


title: 9L2_M_1219


title: 9L2_M_1222


title: 9L2_M_1218


title: 9L2_M_1224


title: 9L2_M_1243


title: 9L2_M_1217


title: 9L2_M_1214


title: 9L2_M_1215


title: 9L2_M_1236


title: 9L2_M_1213


title: 9L2_M_1211


title: 9L2_M_1210


title: 9L2_M_1207


title: 9L2_M_1212


title: 9L2_M_1208


title: 9L2_M_1206


title: 9L2_M_1209


title: 9L2_M_1204


title: 9L2_M_1220


title: 9L2_M_1205


title: 9L2_M_1200


title: 9L2_M_1202


title: 9L2_M_1199


title: 9L2_M_1198


title: 9L2_M_1216


title: 9L2_M_1201


title: 9L2_M_1195


title: 9L2_M_1194


title: 9L2_M_1192


title: 9L2_M_1196


title: 9L2_M_1197


title: 9L2_M_1193


title: 9L2_M_1191


title: 9L2_M_1190


title: 9L2_M_1188


title: 9L2_M_1185


title: 9L2_M_1184


title: 9L2_M_1187


title: 9L2_M_1183


title: 9L2_M_1189


title: 9L2_M_1203


title: 9L2_M_1180


title: 9L2_M_1181


title: 9L2_M_1177


title: 9L2_M_1179


title: 9L2_M_1176


title: 9L2_M_1182


title: 9L2_M_1178


title: 9L2_M_1175


title: 9L2_M_1172


title: 9L2_M_1171


title: 9L2_M_1174


title: 9L2_M_1170


title: 9L2_M_1173


title: 9L2_M_1169


title: 9L2_M_1167


title: 9L2_M_1166


title: 9L2_M_1165


title: 9L2_M_1163


title: 9L2_M_1162


title: 9L2_M_1161


title: 9L2_M_1160


title: 9L2_M_1158


title: 9L2_M_1156


title: 9L2_M_1155


title: 9L2_M_1154


title: 9L2_M_1153


title: 9L2_M_1150


title: 9L2_M_1151


title: 9L2_M_1168


title: 9L2_M_1149


title: 9L2_M_1146


title: 9L2_M_1147


title: 9L2_M_1148


title: 9L2_M_1139


title: 9L2_M_1135


title: 9L2_M_1164


title: 9L2_M_1140


title: 9L2_M_1145


title: 9L2_M_1136


title: 9L2_M_1138


title: 9L2_M_1134


title: 9L2_M_1143


title: 9L2_M_1142


title: 9L2_M_1159


title: 9L2_M_1137


title: 9L2_M_1144


title: 9L2_M_1157


title: 9L2_M_1130


title: 9L2_M_1132


title: 9L2_M_1129


title: 9L2_M_1128


title: 9L2_M_1127


title: 9L2_M_1152


title: 9L2_M_1126


title: 9L2_M_1124


title: 9L2_M_1123


title: 9L2_M_1122


title: 9L2_M_1117


title: 9L2_M_1120


title: 9L2_M_1121


title: 9L2_M_1119


title: 9L2_M_1133


title: 9L2_M_1112


title: 9L2_M_1113


title: 9L2_M_1110


title: 9L2_M_1109


title: 9L2_M_1115


title: 9L2_M_1106


title: 9L2_M_1104


title: 9L2_M_1114


title: 9L2_M_1107


title: 9L2_M_1100


title: 9L2_M_1111


title: 9L2_M_1102


title: 9L2_M_1125


title: 9L2_M_1108


title: 9L2_M_1099


title: 9L2_M_1096


title: 9L2_M_1097


title: 9L2_M_1093


title: 9L2_M_1095


title: 9L2_M_1101


title: 9L2_M_1092


title: 9L2_M_1090


title: 9L2_M_1091


title: 9L2_M_1089


title: 9L2_M_1085


title: 9L2_M_1088


title: 9L2_M_1084


title: 9L2_M_1086


title: 9L2_M_1083


title: 9L2_M_1087


title: 9L2_M_1103


title: 9L2_M_1082


title: 9L2_M_1081


title: 9L2_M_1076


title: 9L2_M_1079


title: 9L2_M_1077


title: 9L2_M_1078


title: 9L2_M_1074


title: 9L2_M_1080


title: 9L2_M_1072


title: 9L2_M_1070


title: 9L2_M_1071


title: 9L2_M_1068


title: 9L2_M_1064


title: 9L2_M_1065


title: 9L2_M_1069


title: 9L2_M_1073


title: 9L2_M_1063


title: 9L2_M_1060


title: 9L2_M_1066


title: 9L2_M_1059


title: 9L2_M_1075


title: 9L2_M_1056


title: 9L2_M_1057


title: 9L2_M_1053


title: 9L2_M_1055


title: 9L2_M_1054


title: 9L2_M_1052


title: 9L2_M_1058


title: 9L2_M_1050


title: 9L2_M_1048


title: 9L2_M_1047


title: 9L2_M_1046


title: 9L2_M_1049


title: 9L2_M_1044


title: 9L2_M_1045


title: 9L2_M_1043


title: 9L2_M_1061


title: 9L2_M_1062


title: 9L2_M_1040


title: 9L2_M_1042


title: 9L2_M_1041


title: 9L2_M_1037


title: 9L2_M_1038


title: 9L2_M_1036


title: 9L2_M_1035


title: 9L2_M_1034


title: 9L2_M_1051


title: 9L2_M_1032


title: 9L2_M_1029


title: 9L2_M_1030


title: 9L2_M_1027


title: 9L2_M_1067


title: 9L2_M_1026


title: 9L2_M_1024


title: 9L2_M_1025


title: 9L2_M_1022


title: 9L2_M_1019


title: 9L2_M_1018


title: 9L2_M_1021


title: 9L2_M_1023


title: 9L2_M_1039


title: 9L2_M_1016


title: 9L2_M_1015


title: 9L2_M_1011


title: 9L2_M_1033


title: 9L2_M_1012


title: 9L2_M_1013


title: 9L2_M_1014


title: 9L2_M_1010


title: 9L2_M_1008


title: 9L2_M_1003


title: 9L2_M_1005


title: 9L2_M_1007


title: 9L2_M_1020


title: 9L2_M_1017


title: 9L2_M_1000


title: 9L2_M_1031


title: 9L2_M_996


title: 9L2_M_994


title: 9L2_M_993


title: 9L2_M_1028


title: 9L2_M_1006


title: 9L2_M_992


title: 9L2_M_990


title: 9L2_M_1004


title: 9L2_M_995


title: 9L2_M_1001


title: 9L2_M_1009


title: 9L2_M_1002


title: 9L2_M_997


title: 9L2_M_991


title: 9L2_M_987


title: 9L2_M_989


title: 9L2_M_986


title: 9L2_M_998


title: 9L2_M_988


title: 9L2_M_985


title: 9L2_M_980


title: 9L2_M_981


title: 9L2_M_982


title: 9L2_M_977


title: 9L2_M_979


title: 9L2_M_999


title: 9L2_M_983


title: 9L2_M_972


title: 9L2_M_976


title: 9L2_M_970


title: 9L2_M_978


title: 9L2_M_973


title: 9L2_M_974


title: 9L2_M_971


title: 9L2_M_969


title: 9L2_M_966


title: 9L2_M_964


title: 9L2_M_968


title: 9L2_M_965


title: 9L2_M_963


title: 9L2_M_984


title: 9L2_M_962


title: 9L2_M_967


title: 9L2_M_960


title: 9L2_M_959


title: 9L2_M_956


title: 9L2_M_958


title: 9L2_M_975


title: 9L2_M_955


title: 9L2_M_957


title: 9L2_M_954


title: 9L2_M_953


title: 9L2_M_947


title: 9L2_M_950


title: 9L2_M_949


title: 9L2_M_952


title: 9L2_M_945


title: 9L2_M_951


title: 9L2_M_943


title: 9L2_M_942


title: 9L2_M_946


title: 9L2_M_941


title: 9L2_M_939


title: 9L2_M_940


title: 9L2_M_938


title: 9L2_M_936


title: 9L2_M_944


title: 9L2_M_935


title: 9L2_M_931


title: 9L2_M_932


title: 9L2_M_933


title: 9L2_M_929


title: 9L2_M_925


title: 9L2_M_928


title: 9L2_M_927


title: 9L2_M_926


title: 9L2_M_922


title: 9L2_M_924


title: 9L2_M_921


title: 9L2_M_920


title: 9L2_M_919


title: 9L2_M_918


title: 9L2_M_923


title: 9L2_M_916


title: 9L2_M_915


title: 9L2_M_914


title: 9L2_M_917


title: 9L2_M_913


title: 9L2_M_934


title: 9L2_M_909


title: 9L2_M_937


title: 9L2_M_908


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