Illumina Genome Analyzer II sequencing; ChIP-Seq for RIP140, using the human breast cancer cell line MCF-7 in the absence of hormone or after treatment with Estradiol
ChIP-Seq for RIP140, using the human breast cancer cell line MCF-7 in the absence of hormone or after treatment with Estradiol
MCF7 cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics. For hormone depletion, cells were grown in phenol red-free DMEM with 5% charcoal-treated FBS and 1% antibiotics for 3 days. Hormone-deprived cells were treated for 3 hours with 10nM Estradiol, dissolved in 0.01% DMSO Cells were fixed with 1% PFA for 10 minutes and processed as described (Schmidt et al., Methods 48:3 pp240-248 2009). For each ChIP, 10ug of antibody was used, and 100ul of Protein A magnetic beads (Invitrogen). The antibody used was raised to detect RIP140 (6D7). ChIP was performed and libaries were contructed as described (Schmidt et al., Methods 48:3 pp240-248 2009).
Experimental Factor: compound
estradiol
Experimental Factor: dose
10 nanomolar
LIBRARY_STRAND
first strand