Figure 7.

Down-regulation of FKBP51 predominantly inhibits androgen-dependent growth in PCa cells. AD LNCaP and AI C4-2 cells were transiently transfected with FKBP51-siRNA or control-siRNA. Six to 8 h after transfection, medium was changed. Twenty-four hours after transfection, cells were plated (1500 cells / well) in 96-well (cell proliferation assay) and 6-well plates (Western blotting). Seventy-two hours after transfection, LNCaP (a) and C4-2 (c) cells were treated with R1881 (0.1 and 1.0 nM) or vehiclel (0) for 24 h. Whole cell extracts were prepared, and expression of Cyp40 and actin (control) was measured by Western blotting. LNCaP (b) and C4-2 (d) cells were treated with R1881 (0.1 and 1.0 nM) or vehicle (0) alone for 3 days. Ninety-six hours after transfection, cell proliferation was determined by means of an MTT assay. Results are means ± SEM of two independent experiments, each performed in quadruplicate. LNCAP: pair-wise comparison of control siRNA vs FKBP51 siRNA at each condition, ***P<0.001. C4-2: pair-wise comparison of control siRNA vs FKBP51 siRNA at each condition, ***P<0.001, **P<0.01,*P<0.05; comparison of control siRNA in presence and absence of R1881, ^##P<0.01.