RNA-sequencing of human breast cancer cell lines treated and untreated with progesterone
RNA-sequencing of human breast cancer cell lines treated and untreated with progesterone
Each sample was collected by scrapping the cells from a 10cm cell culture dish. Pellet was stored at -20 degrees. T47D and MDA-MB-231 cells were cultured in RPMI and DMEM medium respectively. The media was supplemented with 10% FBS, 2.5 mg/ml Amphotericin-B (Abbott) and 1.25 μl/ml Gentamycin (Abbott). The cells were cultured at 37 °C in a 5% CO2 incubator. Cells were serum starved followed by treatment with 10nM of progesterone for 6 hours. Equal volume of ethanol was used as vehicle control RNA was extracted using the TRIzol solubilization and extraction protocol, as recommended by the manufacturer. Transcriptome sequencing libraries were constructed by using the TruSeq RNA library protocol (Illumina). Briefly, ribosomal RNA was depleted and library preparation was done as per the manufacturer's instructions (TruSeq RNA Sample Preparation Kit, Illumina).