FIG. 4.

Pin1 represses β-catenin coactivation of the AR LBD but does not repress CPA-liganded AR. (A) CV1 cells were transfected with pBIND-AR-LBD (50 ng), pG5-Luciferase (10 ng), β-catenin, and Pin1 vectors as indicated (−, not used). Luciferase activities were determined 24 h after DHT treatment. (B and C) CV1 cells were transfected with AR DBD-LBD (50 ng) (B) or AR N-DBD (30 ng) (C) vectors, ARE₄-luciferase reporter (10 ng), β-catenin, and Pin1 as indicated. (D and E) CV1 cells were transfected with pCIneo-AR (10 ng), ARE₄-luciferase reporter (10 ng), and β-catenin and Pin1 expression vectors as indicated. Transfected cells were then treated for 24 h with DHT (D) or CPA (E). pRL-CMV (2.5 ng) was used as an internal control. (F) CV1 cells were transfected as above with pBIND-AR-LBD (50 ng), pG5-Luciferase (10 ng), β-catenin (50 ng), and wild-type (WT) or K63A (KA) mutant Pin1. The percent inhibition (inhib) of control (no Pin1) activity is shown. Pin1 immunoblots were carried out on pooled protein from the triplicate samples. For panels A to E, results are given in relative light units (RLU).