Fig. 4
CWP232291 downregulates the androgen receptor and its splice variants. a AR status in prostate cancer cell lines. Western blot analysis was performed using AR, AR-Vs, and AR-V7 antibodies. GAPDH was used as a loading control. b Cells were exposed to 0–10 μM CWP232291 for 72 h. Cell viability was determined using CellTiter Glo® (means ± SD, n = 3). c LNCaP and 22Rv1 cells were exposed to CWP232291 for 72 h at the IC50. Western blotting was performed using AR antibodies. Actin was used as a loading control. d LNCaP and 22Rv1 cells were exposed to CWP232291 for 24 h at the IC50 and then stained with AR (red) and DAPI (blue). Images were captured using a fluorescence microscope. Scale bar, 100 μm. e LNCaP and (F) 22Rv1 cells were androgen-deprived for 48 h and then treated with vehicle or 1 nM DHT with or without IC50 doses of CWP (LNCaP 100 nM; 22Rv1 60 nM) for 24 h. Relative mRNA levels of AR, PSA, UBE2C, and TMPRSS2 were quantified by real-time PCR in prostate cancer cells