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Status |
Public on Apr 17, 2024 |
Title |
Naive, rep2 |
Sample type |
SRA |
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Source name |
B cells
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Organism |
Mus musculus |
Characteristics |
strain: C57BL/6 cell type: B cells genotype: WT
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Extracted molecule |
genomic DNA |
Extraction protocol |
For B cell isolation, splenic B cells were purified by the negative selection of CD43+ cells with anti-CD43 magnetic beads (Militenyi Biotec). The purified B cell population was>95% positive for CD19 staining. Cell sorting was performed on FACSMelody (BD Biosciences) to isolate the following populations: activated B cells (CD138lowB220hi) ; plasma cells (CD138hiB220low) and retrovirus-infected cells (GFP+) .For B cell stimulation assays, purified B cells were cultured in RPMI 1640 medium. For plasma cell differentiation, B cells were stimulated with 5 μg/ml of lipopolysaccharide (LPS). Assay for transposase-accessible chromatin sequencing (ATAC-seq): ATAC-seq was performed following published protocol (Corces, M. et al., Nat Methods, 2017). 50,000 cells were used for each library. Chromatin integration labelling (ChIL): ChIL-seq for B cells was performed described previously (https://doi.org/10.1038/protex.2018.122), except following. Cells were biotinylated for 30 min at 4°C with 1mg/ml sulfoNHS-SS-biotin in PBS and plated on streptavidin-treated 96-well plates (Thermo Fisher Scientific). After fixed with 1% paraformaldehyde (Electron Microscopy Sciences) in PBS for 5min at room temperature, fixation was stopped with 1 M Glycine/HCl.
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Library strategy |
ATAC-seq |
Library source |
genomic |
Library selection |
other |
Instrument model |
Illumina NovaSeq 6000 |
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Description |
B cells
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Data processing |
After sequencing, reads were cleaned by trim_galore (version0.6.10) and mapped to mm10 with Bowtie2(version 2.3.1). Replicate samples were merged and peaks were identified by using MACS2(ChIL: -q 0.01 --nomodel –nolambda, ATAC: -q 0.01) Bigwig files were prepared using deeptools with following options "--binSize 100 --centerReads --normalizeUsing CPM" for ATAC-seq and "--smoothLength 1000 --binSize 100 --normalizeUsing CPM" for ChIL-seq. Assembly: mm10 Supplementary files format and content: peak summit files(.bed), bigwig files(.bigwig)
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Submission date |
Apr 24, 2023 |
Last update date |
Apr 17, 2024 |
Contact name |
Yasuyuki Ohkawa |
E-mail(s) |
yohkawa@bioreg.kyushu-u.ac.jp
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Organization name |
Medical Institute of Bioregulation
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Lab |
Division of Transcriptomics
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Street address |
3-1-1 Maidashi
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City |
Fukuoka |
ZIP/Postal code |
8128582 |
Country |
Japan |
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Platform ID |
GPL24247 |
Series (1) |
GSE230496 |
Epigenomic profiling by ATAC-seq and ChIL-seq in B cells and plasma cells |
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Relations |
BioSample |
SAMN34357486 |
SRA |
SRX20089811 |
Supplementary file |
Size |
Download |
File type/resource |
GSM7225370_20220908_ATAC_Naive_Cr0727_1_Ad3.bin100PE.CPM.bw |
33.1 Mb |
(ftp)(http) |
BW |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
Processed data are available on Series record |
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