Lncap 1f5

The LNCaP, 22Rv1, PC3, VCaP, RWPE1, RWPE2 and 293 T cell lines were originally obtained from the American Type Culture Collection (ATCC), and have been authenticated by STR profiling. The LNCaP-1F5 and V16A cell lines were kindly provided by Prof. Hansen He, University of Toronto and Prof. Olli A Jänne, University of Helsinki, respectively. All cell lines were tested regularly for mycoplasma (EZ-PCR Mycoplasma Test Kit, 20–700-20, Biological Industries). All cell lines were found to be negative of mycoplasma during our study. LNCaP, LNCaP-1F5, V16A, PC3 and 22Rv1 cells were cultured in RPMI1640 medium (Sigma), VCaP and 293T cells were grown in DMEM (Invitrogen), RWPE1 and RWPE2 cells were cultured in Keratinocyte-Serum Free Medium (KSF, Invitrogen). All mediums were supplemented with 10% FBS (Gibco) and 1% antibiotics (penicillin and streptomycine, Sigma), with the exception of KSF medium that was supplemented with the epidermal growth factor (EGF) and Bovine Pituitary Extract (BPE) included in the kit (17,005–042, Introvigen). To induce androgen signaling and AR activity, relevant PCa cells were treated with dihydrotestosterone (DHT, Sigma) for indicated time.

RWPE1, DU145, PC3, 22Rv1, LNCaP, VCaP and JIMT-1 cell lines were purchased from ATCC. LNCaP 1F5 and V16A were a gift from Dr. Olli Jänne from the University of Helsinki. RWPE1 cells were cultured in Keratinocyte SFM medium (Gibco) supplemented with bovine pituitary extract and human recombinant EGF, and standard antibiotics: penicillin (100 units/ml) and streptomycin (100 μg/ml), according to the manufacturer’s protocol. PC3 and DU145 cells were maintained in F12K (Gibco) and MEM (Gibco) medium, respectively containing 10% fetal bovine serum (Gibco) and standard antibiotics. 22Rv1, VCap, LNCaP, LNCaP 1F5, V16A and JIMT-1 cells were grown in RPMI1640 (Sigma-Aldrich) containing 10% FBS and standard antibiotics. The cells were maintained at 37 °C in a humidified atmosphere with 5% CO₂. All cell lines were confirmed to be mycoplasma-free during the analysis.