Transcription profiling by array of human MCF-7 TET Off breast cancer cells (MCF-7 wt) to investigate unliganded Estrogen Receptor Beta on miRNome downstream target regulation

MCF-7 TET Off cells (MCF-7 wt) were used to produce stable clones expressing ER-beta tagged with TAP-tag at the C-term (C-TAP-ER-beta). All cells were grown in Dulbecco's modified Eagle's medium (DMEM), starved by using DMEM w/o phenol red and 5% Dextran Coated Charcoal stripped serum (DCC-FBS) for 5 days. Cells were transfected wiyh hsa-miR-30a-5p Mimics and with tha negative control Scramble both at concentration of 5 and 10 nM. Biological replicate were lysed and RNA extracted were pooled 24 hrs after transfectiions. For mRNA expression profiling, 500 ng total RNA were reverse transcribed and used for synthesis of cDNA and biotinylated cRNA. Finally cRNA were hybridized for 19 hours on Illumina HumanHT-12 v4.0 BeadChips and after scanning, data analysis was performed.