The overexpression of wild-type or mutant AR potentiates Wnt signaling. Cells grown in RPMI with 10% FBS were transiently transfected with indicated plasmids together with LEF-luciferase and pCMV-Renilla reporter constructs in 12 well plates. Cotransfection of Wnt1 and AR leads to a synergistic effect on the activation of LEF-luciferase reporter in PC3 cells (A), CWR22Rv1 and LNCaP cells (B), compared with transfection with Wnt1 alone. The AR can also potentiate the activation of Wnt signaling in cells expressing a constitutively activated human receptor LRP6, truncated at the N terminus (H6ΔN), or a stabilized β-Catenin S37A (C). The synergistic effect between AR mutants and Wnt1 signaling is shown (D). AR expression levels were measured for both wild type and mutant ARH874Y. Actin levels were used as loading controls. The effect of an AR antagonist is shown (E & F). PC3 cells were treated with bicalutamide 4 hours after transfection. Dual luciferase assay was performed 20–24 hours after treatment.