NextSeq 500 paired end sequencing; RNA-Seq of human breast cancer cells MCF7 treated with EPZ004777 or DMSO for 3 or 6 days
RNA-Seq of human breast cancer cells MCF7 treated with EPZ004777 or DMSO for 3 or 6 days
ER-alpha positive BC cells: MCF-7 (ATCC HTB-22), were propagated in Dulbecco's modified Eagle medium (DMEM; Sigma-Aldrich) supplemented with 10% FBS (HyClone) and antibiotics: 100 U/ml penicillin, 100 mg/ml streptomycin, 250 ng/ml amphotericin-B.  None ER-alpha positive BC cells: MCF-7 (ATCC HTB-22) were propagated in Dulbecco's modified Eagle medium (DMEM; Sigma-Aldrich) supplemented with 10% FBS (HyClone) and antibiotics: 100 U/ml penicillin, 100 mg/ml streptomycin, 250 ng/ml amphotericin-B. 104cells/well were incubated with compounds for 3 or 6 days with media containing compounds or vehicle and in particular MCF-7 cells treated with EPZ004777 6.4µM or DMSO for for 3 and 6 days or with 4-hydroxytamoxifen 10-7 M  (4-OHT) or fulvestrant 10-7 M (ICI 182,780) or DMSO for 3 days. None 104cells/well were incubated with compounds for 3 or 6 days with media containing compounds or vehicle and in particular MCF-7 cells treated with EPZ004777 6.4µM or DMSO for for 3 and 6 days or with 4-hydroxytamoxifen 10-7 M  (4-OHT) or fulvestrant 10-7 M (ICI 182,780) or DMSO for 3 days. For RNA sequencing experiments, indexed libraries were prepared using 1 µg of total RNA from cells with TruSeq Stranded Total RNA Sample Prep Kit (Illumina Inc.).
Experimental Factor: compound
EPZ004777
Experimental Factor: dose
6.4
Experimental Factor: time
3