Full bovine serum fbs

Most cell lines were originally purchased from the American Type Culture Collection (ATCC) and were cultured as per the standard ATCC protocols. LNCaP-AR and LAPC4 cells were gifts from Dr. Charles Sawyers lab (Memorial Sloan-Kettering Cancer Center, New York, NY). Until otherwise stated, for all the experiments LNCaP, PNT2, LNCaP-AR, C42B, 22RV1, DU145, PC3 cells were grown in the RPMI 1640 medium (Gibco) and VCaP cells in the DMEM with Glutamax (Gibco) medium supplemented with 10% Full Bovine Serum (FBS; Invitrogen). LAPC4 cells were grown in IMEM (Gibco) medium supplemented with 15%FBS and 1nM of R1881. Immortalized normal prostate cells: RWPE1 were grown in keratinocyte media with regular supplements (Lonza); PNT2 were grown in RPMI medium with 10%FBS. HEK293 cells were grown in DMEM (Gibco) medium with 10% FBS. All cells were grown in a humidified 5%CO2 incubator at 37℃. All cell lines were biweekly tested to be free of mycoplasma contamination and genotyped every month at the University of Michigan Sequencing Core using Profiler Plus (Applied Biosystems) and compared with corresponding short tandem repeat (STR) profiles in the ATCC database to authenticate their identity in culture between passages and experiments.