(A to C) C4–2B cells were retrovirally infected with vector, AR, or K609A-AR constructs. After 96 hours, RNA was extracted, followed by qRT-PCR using primers corresponding to AR (A), KLK3 (B), or TMPRSS2 (C). (D to G) 22Rv1, MCF7, and LNCaP cells were transfected with vector, AR, and K609A-AR constructs. Forty-eight hours after transfection, RNA was isolated, followed by qRT-PCR using primers corresponding to PSA [(D) 22Rv1; (E) MCF7] or NRG3 [(F) MCF7; (G) LNCaP]. (H to J) VCaP cells were treated overnight with (R)-9b (5 μM), SBHA (1 μM), or C646 (5 μM). RNA was isolated, followed by qRT-PCR using primers corresponding to AR (H), PDCD6IP (I), and KLK3 (J). (K to M) HEK293 (K), PC3 (L), and MCF7 (M) cells were transfected with AR, K609A-AR mutant, or K609Q-AR mutant and the ARR2PB-luciferase reporter construct. Cells were treated with or without 10 nM DHT overnight in serum-free media, and luciferase activity was determined 48 hours after transfection. For (A to M), n = 2, three replicates; representative data are shown. Data are represented as means ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, Student’s t test.