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. 2004 Jul;24(13):6021–6028. doi: 10.1128/MCB.24.13.6021-6028.2004

FIG. 1.

FIG. 1.

SENP1 markedly enhances AR-dependent transcription. (A) Enhancement of AR-dependent transcription by SENP1, but not by mutant SENP1. PC-3 cells were transfected with AR (10 ng) and ARE-luciferase (50 ng) reporter plasmids in the absence or presence of wild-type or mutant SENP1 plasmids (150 ng). After 12 h of transfection, cells were treated with 10 nM R1881 for 24 h, and the luciferase activity was measured. Transfection efficiency was normalized with a β-galactosidase expression construct, and the results are presented as activation over that for an empty vector. The expression level of AR was analyzed by Western blotting with the anti-AR antibody. (B) Dose response of SENP1 action. PC-3 cells were transfected with AR (10 ng) and ARE-luciferase (50 ng) reporter plasmids in the absence or presence of increasing amounts of wild-type or mutant SENP1 plasmids (10, 50, and 150 ng). After 12 h of transfection, cells were treated with 10 nM R1881 for 24 h, and the luciferase activity was measured as described for panel A. (C) SENP1 could not activate antagonist-bound AR. PC-3 cells were transfected with AR (10 ng) and ARE-luciferase (50 ng) reporter plasmids in the absence or presence of wild-type or mutant SENP1 plasmids (150 ng). Twelve hours after transfection, cells were treated with 10 nM R1881 or 5 μM bicalutamide for 24 h, and the luciferase activity was measured as described for panel A. (D) SENP1, but not SENP2 or SENP3, markedly activates AR transactivation. PC-3 cells were transfected with AR (10 ng) and ARE-luciferase (50 ng) reporter plasmids in the absence or presence of SENP1, SENP2, or SENP3 plasmids (150 ng). Twelvehours after transfection, cells were treated with 10 nM R1881 for 24 h, and the luciferase activity was measured as described in for panel A. (E and F) Western blots of cell extracts from panels B (E) and D (F). (G) SENP1 enhances probasin promoter activity. PC-3 cells were transfected with AR (10 ng) and ARE-luciferase or PB(−426/+28)-luciferase (50 ng) reporter plasmids in the absence or presence of wild-type or mutant SENP1 plasmids (150 ng). After 12 h of transfection, cells were treated with 10 nM R1881 for 24 h, and the luciferase activity was measured as described for panel A.