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Sample GSM1164146 Query DataSets for GSM1164146
Status Public on Jun 02, 2014
Title FOXA1_ChIP-seq_DU145 AR + FoxA1 high
Sample type SRA
 
Source name FOXA1_ChIP-seq_DU145 AR + FoxA1 high
Organism Homo sapiens
Characteristics cell line: prostate cancer cell line DU145
ectopic expression: AR wildtype + FoxA1 high
chip antibody: Anti-FOXA1 antibody (Abcam, cat#ab23738,lot# GR77830-1)
Treatment protocol DU145 AR cells were infected with LacZ or FOXA1 adenovirus for 48 hours to overexpress FOXA1 protein prior to collect cell pellets for ChIP or FAIRE experiments
Growth protocol DU145 cells were routinely maintained in RPMI supplemented with 10% FBS plus 1% antibiotics (penicillin and streptomycin)
Extracted molecule genomic DNA
Extraction protocol Antibodies were from abcam (cat#ab23738,GR77830-1) and Millipore (cat#06-680, lot#JBC1939961) and ChIP was performed using 5e6-10e6 cells per reaction. Chromatin was fixed in room temperature for 10 minutes with serum free media containing 0,37 % formaldehyde. Cells were washed twice in PBS and scraped before treatment of cell lysis buffer with protease inhibitors on ice. Pelleted nuclei were sheared to 200-700bp using an ultrasonic sonicator (Misonix). ChIP was done with 5 micrograms of antibody. FAIRE analysis was performed according to the protocol published by Giresi et al .
Libraries were prepared according to Bioo Scientific's instructions accompanying the DNA Sample Kit (Part# 514101). 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 (3' to 5' 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 Illumina HiSeq 2000 or Illumina HiSeq 2500 following the manufacturer's protocols.
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina HiSeq 2000
 
Description ChIP-seq
2012-833
Data processing Basecalls performed using CASAVA version 1.4
ChIP-seq reads were aligned to the hg19 genome assembly using bwa version 0.61
Peak-calling by HOMER v4.1
Genome_build: hg19
Supplementary_files_format_and_content: Columns in peaks.txt contain information about each peak: Column 1: PeakID - a unique name for each peak. Column 2: chr - chromosome where peak is located. Column 3: starting position of peak. Column 4: ending position of peak. Column 5: Strand (+/-). Column 6: Normalized Tag Counts - number of tags found at the peak, normalized to 10 million total mapped tags (or defined by the user). Column 7: Focus Ratio - fraction of tags found appropriately upstream and downstream of the peak center (see below). Column 8: Peak score (position adjusted reads from initial peak region - reads per position may be limited). Columns 9+: Statistics and Data from filtering.
 
Submission date Jun 15, 2013
Last update date May 15, 2019
Contact name Hong-Jian Jin
E-mail(s) hongjian-jin@northwestern.edu
Phone 3125033041
Organization name Northwestern University
Department Medicine
Street address 303 E Superior St
City Chicago
State/province IL - Illinois
ZIP/Postal code 60611
Country USA
 
Platform ID GPL11154
Series (2)
GSE47987 FoxA1 inhibits androgen receptor expression and suppresses prostate cancer metastasis [DU145, ChIP-seq]
GSE55007 Cooperativity and Equilibrium with FOXA1 Define Androgen Receptor Transcriptional Program
Relations
BioSample SAMN02204346
SRA SRX306511

Supplementary file Size Download File type/resource
GSM1164146_2012-833.peaks.txt.gz 3.8 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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