Illumina HiSeq 2500 sequencing; Chip-Seq of wild-type GATA3 and two mutants (335fs, 308*) in the ZR75-1 human breast cancer cell line
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
Breast
Cell type
ZR-75-1
Primary Tissue
Breast
Tissue Diagnosis
Adenocarcinoma Ductal
Attributes by original data submitter
Sample
ENA first public
2019-01-18
ENA last update
2019-01-08
ENA-CHECKLIST
ERC000011
External Id
SAMEA5207806
INSDC center alias
Kabara Cancer Research Institute
INSDC center name
Kabara Cancer Research Institute
INSDC first public
2019-01-18T17:02:25Z
INSDC last update
2019-01-08T17:31:39Z
INSDC status
public
Submitter Id
E-MTAB-7578:308_FLAG
broker name
ArrayExpress
cell line
ZR-75-1
common name
human
developmental stage
adult
disease
invasive ductal carcinoma
individual
GATA3 308*
organism part
mammary gland
sample name
E-MTAB-7578:308_FLAG
sex
female
Sequenced DNA Library
library_name
308_FLAG_s
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Approx 8 million adherent cells were crosslinked for 10 mins in 1% formaldehyde. Cross linking was stopped by adding glycine to 0.125M. Cells were lysed in Farnham Lysis Buffer, washed in PBS and nuclei were lysed with RIPA buffer. ZR75-1 cells were cultured in RPMI 1640 supplemented with 10% serum Cell lines were generated by retroviral transduction of ZR75-1 with vectors containing either GATA3 WT, 308 or 335 mutations. Samples were sonicated using a Diagenode XL Bioruptor to fragments of 200-400 bp. Chromatin immunoprecipitation was performed using anti-FLAG antibody bound to protein G magnetic dynabeads. DNA fragments recovered from chromatin immunoprecipitation were reapired using Klenow DNA polymerase, liganted to Illumina TruSeq adapters using T4 DNA ligase and size selected for DNA with adapters attached.