Cwr22v1

RWPE1, LNCaP, PC3, C4-2B, DU145, CWR22V1, and HEK293 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA). C4-2B cells resistant to enzalutamide (C4-2B-MDVR) were generated by culturing C4-2B cells to increasing concentrations of enzalutamide (5–40 µM) in media for over 12 months and maintained in complete media supplemented with 20 µM enzalutamide as described previously [58 (link), 59 (link)]. HEK293 cells were cultured in complete DMEM medium. RWPE1, LNCaP, LNCaP-KDM8 [17 (link)], PC3, C4-2B, DU145, and CWR22RV1 cells were cultured in RPMI-1640 medium supplied with 10% heat-inactivated FBS, 100 U/ml penicillin, and 0.1 mg/ml streptomycin in a humidified incubator with 5% CO₂. For hormone induction experiments, the growth media were replaced by RPMI-1640 containing 10% charcoal-dextran-treated (CDT) FBS (CDT-FBS) for 2 days before treatment with 10 nM dihydrotestosterone (DHT).

Luciferase-bearing PC-3M and PC-3S cells were clonally derived from the human cell line PC-3 [11 (link)]. Du-145, CWR22v1 and LNCaP cells were obtained from the American Type Culture Collection (Manassas, VA). Cells were grown at 37°C in a 5% CO₂ atmosphere in RPMI 1640 medium supplemented with 10% fetal bovine serum, L-glutamine, non-essential aminoacids, sodium pyruvate, penicillin/streptomycin (PAA Laboratories, Coelbe, Germany) and geneticin (Santa Cruz Biotechnologies, Santa Cruz, CA).

Anti-FLAG antibody was purchased from eBioscience (Cat#14-6681-82), anti-panCK antibody
from Biogenex (Cat#AM273-5 M), anti-VHL antibody from Abcam (Cat#ab140989), and
anti-CK8/18 from Novus (Cat#NBP2-44929). Murine ccRCC cell line RENCA and human ccRCC cell
line ACHN, prostate cancer cell lines CWR22v1 and C4-2 were purchased from the American
Type Culture Collection (ATCC) and maintained in RPMI-1640, supplemented with 10% fetal
bovine serum and penicillin/Streptomycin at a working concentration of 100 U/mL. Murine
prostate cancer cells Myc-CaP were purchased from ATCC and maintained in DMEM,
supplemented with 10% fetal bovine serum and penicillin/Streptomycin at a working
concentration of 100 U/mL. Human bladder cancer cell lines T24 and HT-1376 were kind gifts
from Dr. Arnold I. Chin and Dr. Hanwei Zhang at UCLA and maintained in the same condition
as RENCA cells.
Lentiviral plasmid encoding mStrawberry and EGFP, together with flag tag or HA, and
plasmid encoding firefly luciferase were constructed based on pSicoR (Addgene, #11579),
and lentivirus was packaged as mentioned previously in the report²⁶ (link).