ProFine inhibits AR and survivin signaling in PCa cells. (A) RT-PCR (upper, 24 hours treatment) and Western blot analyses (bottom, 11.6 μg/ml) of AR expression in C4-2 cells treated with ProFine. (B) Upper: Western blot analysis of AR protein expression in C4-2 cells pretreated with CHX (50 μg/ml, 1 hour) and further treated with ProFine (11.6 μg/ml) or DMSO for the indicated times. Lower: Plot of AR protein degradation in C4-2 cells treated with ProFine or DMSO. (C) Western blot analysis of AR protein expression in HSP90 immunoprecipitates from C4-2 cells treated with ProFine (11.6 μg/ml, 24 hours) or DMSO. (D) RT-PCR analysis of survivin expression in C4-2 cells treated with ProFine (24 hours); (E) Western blot analysis of p-Stat3, Stat3, survivin, and PARP in C4-2 cells treated with ProFine (11.6 μg/ml) at indicated times. (F) Upper: Schematic diagram of the Stat3 cis-elements in human survivin promoter; lower: luciferase activity of human survivin promoters in C4-2 cells treated with ProFine (11.6 μg/ml). (G) Western blot analysis of protein expression of AR, HIF-1α, and VEGF in C4-2 cells treated with ProFine (11.6 μg/ml). (H) In vitro tube formation of HUVECs treated with varying concentrations of ProFine (72 hours).