organism: Homo sapiens
description: MCF-7 parental cells were stably transduced with a doxycycline-inducible mEmerald vector (MCF7-EM) or ESR1-mEmerald vector (MCF7-ER) and maintained in media containing 10% Tet-free FBS (Takara Bio USA, Inc.), 500g/ml Geneticin (Thermo Fisher Scientific) and 1g/ml puromycin (Millipore Sigma). Prior to E2 treatment, cells were rinsed with PBS then adapted in phenol red-free media (Thermo Fisher Scientific) containing 10% carbon-stripped FBS (Millipore Sigma) for three days. After adaptation, the MCF7-EM and MCF7-ER cells were induced with doxycycline for 24 hours, then treated with 10 nM estradiol or vehicle control (100% ethanol) for 1 hour. Treated cells crosslinked with 1% formaldehyde, sonicated, then incubated with ER antibody (HC-20, Santa Cruz-discontinued) overnight. ER-immunoprecipitates were collected with 1.5 mg protein A Dynabeads (Thermo Fisher Scientific), then reverse-crosslinked and digested with 40g/ml proteinase K (Thermo Fisher Scientific). DNA was purified with the ThermoFisher PCR purification Kit (K0702). A 1% input sample was collected for each ChIP experiment to serve as control.
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: age
attributes.value: not applicable
attributes.tag: biomaterial_provider
attributes.value: Judith Hugh, University of Alberta, 5-108 Katz Group Centre for Pharmacy and Health Research
attributes.tag: sex
attributes.value: female
attributes.tag: tissue
attributes.value: mammary gland
attributes.tag: cell_line
attributes.value: MCF-7
attributes.tag: cell_type
attributes.value: epithelial cell
attributes.tag: disease
attributes.value: invasive ductal carcinoma
attributes.tag: treatment
attributes.value: 10 nM Estradiol
attributes.tag: replicate
attributes.value: EM10_ER-Biological replicate-3
attributes.tag: BioSampleModel
attributes.value: Human