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Sample GSM4443871 Query DataSets for GSM4443871
Status Public on Jul 11, 2020
Title Foxa2-OE_Vector-KrasG12D_ChIP
Sample type SRA
Source name pancreatic adenocarcinoma
Organism Mus musculus
Characteristics strain background: B6/FVB
cell line: KrasG12D
cell type: pancreatic adenocarcinoma cancer Tet-on KrasG12D cell line
genotype/variation: tet-on_KrasG12D;KO_Cdkn2a;KO_Smad4;KO_Trp53
chip antibody: anti-Foxa2 (Cell Signaling, #8186)
Treatment protocol KPCS and KP cell line were treated by Refametinib,PD0325901,MK-2206 2HCl or DMSO for 12h.
To KrasG12D cell line: +KrasG12D means treatment with Dox;-KrasG12D means No Dox add.PDA cell line:+KrasG12D means endogenous KrasG12D expression.
Growth protocol KPCS and KP cell line: DMEM+10%FBS+1%P/S.
To KrasG12D cell line: DMEM+10%FBS+1%P/S+1ug/ml Doxycycline; PDA cell line:DMEM+10%FBS+1%P/S
Extracted molecule genomic DNA
Extraction protocol Briefly, cells were crosslinked at room temperature for 10 min with 1% formaldehyde, washed twice with PBS and sonicated in lysis buffer (50 mM HEPES/KOH pH 7.5, 140 mM NaCl, 0.1% Na-Deoxycholate, 1% Triton X-100, 1 mM EDTA, complete protease inhibitor cocktail). Samples were immunoprecipitated at 4 ºC overnight and washed six times with ChIP wash buffer (20 mM Tris, pH 7.9, 500 mM NaCl, 2 mM EDTA, 1% Triton X-100, 0.1% SDS) and once with TE buffer. Bound DNA was eluted in 1% SDS buffer and reverse-crosslinked for 6 h at 65 ºC
DNA samples were treated sequentially with RNase A and Protease K and then purified with ChIP DNA Clean & Concentrator-5 (Zymo Research, D4014)
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model HiSeq X Ten
Description pool12-Foxa2-3-4-V-no-dox_FKDL202555768-1a-1
Data processing The adapters of paired-end reads were trimmed and quality control checks were carried out using trim-galore (version 0.6.0). Trimmed reads were aligned to mouse genome reference (GRCm38.p6) from GENCODE using bowtie2 (version of which the parameter is “–very-sensitive –X 2000”. Aligned reads stored in SAM format were manipulated to generate bam files using samtools (version 1.9) with parameter is “samtools view –b –h –F 1028 –f 3”. Peaks of ChIP-seq were called using MACS2 (version 2.1.2) using the parameter “macs2 callpeak --verbose 3 -g mm -B -q 1e-3 -f BEDPE”. Analysis of differential peaks intensity was processed using MAnorm (version 1.1.4).
Genome_build: mm10
Supplementary_files_format_and_content: bigwig files were generated using deeptools
Submission date Mar 30, 2020
Last update date Jul 12, 2020
Contact name Yi He
Organization name Tsinghua university
Street address 30 Shuangqing Rd, Haidian Qu, Beijing Shi, China
City Beijing
ZIP/Postal code 100084
Country China
Platform ID GPL21273
Series (2)
GSE134233 Mutant Kras co-opts a progenitor-derived enhancer network to initiate pancreatic tumorigenesis [ChIP-seq]
GSE134236 Mutant Kras co-opts a progenitor-derived enhancer network to initiate pancreatic tumorigenesis
BioSample SAMN14485834
SRA SRX8026864

Supplementary file Size Download File type/resource 125.4 Mb (ftp)(http) BW
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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