Fig. 4.

a 22Rv1 cells were transfected with control si-RNA’s (si-Control), si-AR-V7 (to silence AR-V7), si-ex-7 (to silence flAR) and si-ex-2 (to silence both AR-V7 and flAR). Aliquots were used for western analysis of flAR, AR-V7 and beta actin. b Aliquots of cells transfected with si-Control (1 and 4), si-AR-V7 (2 and 5), si-ex-7 (3 and 6) were seeded, grown in CS-FBS for 24 h and treated with vehicle (1, 2, and 3) or CUDC-101 (4, 5 and 6) for 6 h. Bars ± SE indicate PSA/18S mRNA, with siControl treated with vehicle (lane 1) set at 1. Dotted lined emphasize the difference between 1 (si-Control) and 2 (si-AR-V7), and the lack of difference between 2 (si-AR-V7 treated with vehicle) and 5 (si-AR-V7 treated with CUDC-101). c 22Rv1 cells were seeded and grown for 24 h in CS-FBS. Cells were treated with vehicle, DHT (2 nM), MDV3100 (10 μM), Casodex (10 μM) or CUDC-101 (300 nM) for 24 h. Bars ± SE indicate PSA/18S mRNA with cells treated with DHT set at 1. d Aliquots of the experiment presented in A were seeded and grown in the presence of vehicle or CUDC-101 (300 nM) for 72 h. Bars ± SE indicate OD450 as a marker of cell proliferation. Data are normalized to cells grown in the presence of vehicle and transfected with control siRNA. e. 22Rv1 cells were seeded and grown in CS-FBS for 24 h, and treated with vehicle, MDV3100 (10 μM) or Casodex (10 μM) for 72 h. Bars ± SE indicate OD450. Data are normalized to cells grown under control (vehicle) conditions to 100 %. f. Immunoblot analysis for p21, HER2/NEU, Acetyl-H3, Acetyl alpha tubulin and alpha tubulin. 22Rv1 cells were seeded in CS-FBS for 24 h, and treated with vehicle or CUDC-101 for 6 h. Asterisks denote statistical significance: *P < 0.05, ** P < 0.01, *** P < 0.001