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 24 hours. Total RNA was extracted from treated MCF7-EM and MCF7-ER cells treated using the NucleoSpin RNA extraction kit, which included a DNase digestion step to remove potential contaminating DNA (Macherey-Negel). RNA quality was assessed using the Agilent Bioanalyzer 2100 and all samples had RIN scores from 8.6-10. cDNA libraries were made from 1g total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490) and NEBNext Ultra Directional RNA Library Prep Kit for Illumina (E7420) and indexed for sequencing using NEBNext Multiplex Oligos for Illumina. Libraries were sequenced on Illuminas NextSeq 500 using the NextSeq 500/550 High Output Kit (2x75 cycles).
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: 100% ethanol
attributes.tag: replicate
attributes.value: EM0-Biological replicate-3
attributes.tag: BioSampleModel
attributes.value: Human