|
Status |
Public on Nov 17, 2019 |
Title |
HEPG2_PU1_1 |
Sample type |
SRA |
|
|
Source name |
HEPG2_PU1
|
Organism |
Homo sapiens |
Characteristics |
cell type: HEP-G2 cell type: hepatocellular carcinoma cell line transfected ivt mrna: PU.1 molecule subtype: transposed, nuclear DNA
|
Treatment protocol |
Cells were transfected with the indicated in vitro transcribed mRNA transcripts (PU1-IVT-mRNA, PU1mut-IVT-mRNA, PU1-delA-IVT-mRNA) and cultured for 8 hours prior to lysis procedure.
|
Growth protocol |
CTV-1 and HEP-G2 cells were cultured in RPMI 1640 (Gibco; routinely supplemented with 2 mM L-glutamine (Biochrom), 1 mM sodium pyruvate (Sigma), 50 U/ml penicillin/streptomycin, 0.4x vitamins (Sigma), 1x non-essential amino acids (Sigma), 50 μM b-mercaptoethanol (Gibco)) supplemented with 10% heat inactivated fetal bovine serum; For TALL-1 cells the culture medium was supplemented with 15% heat inactivated fetal bovine serum was used; adherent HEP-G2 cells were detached using Trypsin-EDTA (Gibco)
|
Extracted molecule |
genomic DNA |
Extraction protocol |
Cells were treated with DNase I directly in culture medium prior to nuclear transposition procedure. ATACseq was essentially done as described (Corces et al. 2017).
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|
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Library strategy |
ATAC-seq |
Library source |
genomic |
Library selection |
other |
Instrument model |
NextSeq 550 |
|
|
Description |
Cells were transfected with IVT mRNA and transposition was carried out 8 hours after transfection
|
Data processing |
Raw data were aligned to the human genome (GRCh37/hg19) using bowtie2 in very-sensitive and no-discordant modes, keeping only reads that map to a single unique genomic location for further analysis (MAPQ > 10). Read positions were adjusted to move the ends proximal to the Tn5 binding site (for reads on the positive strand, the start is shifted +4 bp and its partner reads start -5 bp, for reads on the negative strand, the start is shifted -5 bp and its partner reads start +4 bp) using a custom script. Bigwigs of ChIPseq data sets were generated using HOMER v4.9 (Heinz et al, 2010) and a fixed fragment length of 65 bp. For cancer cell lines, we used the Control-FREEC v11.0 program to determine allelic imbalances from low-depth whole genome sequencing data and corrected ChIP-seq read counts accordingly to generate CNV-corrected bigWigs. ATAC-seq peak regions were called by combining two different approaches: The basic peak region set was called using HOMER?s findPeaks program in ?region?? mode using parameters ?-size 150 -minDist 250 -L 2 -fdr 0.00001? to identify regions of variable length by stitching nucleosome-size peaks. To exclude shallow peak regions, only those were kept that overlapped a second peak set that was generated in ??factor?? mode using parameters ?-size 250 -minDist 250 -L 2 -fdr 0.00001? to identify focal peaks. Genome_build: hg19hg19 Supplementary_files_format_and_content: coverage tracks in bigWig format (before and after CNV normalization Supplementary_files_format_and_content: ATACseq peaks (open chromatin regions)
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|
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Submission date |
Mar 25, 2019 |
Last update date |
Nov 19, 2019 |
Contact name |
Michael Rehli |
E-mail(s) |
michael.rehli@klinik.uni-r.de
|
Organization name |
University Hospital Regensburg
|
Department |
Internal Med III
|
Street address |
F.-J.-Strauss-Allee 11
|
City |
Regensburg |
ZIP/Postal code |
93042 |
Country |
Germany |
|
|
Platform ID |
GPL21697 |
Series (2) |
GSE128833 |
The hematopoietic master transcription factor PU.1 requires its interaction with the SWI/SNF remodeler to access chromatin de novo [ATAC-seq] |
GSE128837 |
The hematopoietic master transcription factor PU.1 requires its interaction with the SWI/SNF remodeler to access chromatin de novo |
|
Relations |
BioSample |
SAMN11252769 |
SRA |
SRX5574411 |