Long term inhibition of GSK-3β in C4-2 cells.
(A) Silencing of GSK-3β in C4-2 cells. C4-2 cells were transiently transfected with pKD-GSK-3β-v1 or pKD-NegCon-v1. 48 hours after transfection, 10 nM DHT was added where indicated. After another 24 hours, nuclear extracts were prepared as described in Material and Methods. (B) Long term inhibition of GSK-3β using SB216763. AR-positive C4-2 cells were treated with increasing amounts of SB216763 for 48 hours in the absence of androgens. Cells were lysed and cell lysates were analyzed by Western blotting for AR. β-actin bands served as loading control.