Figure 4.

Effects of pirfenidone (PFD) treatment on androgen receptor signaling-related protein levels in human prostate cancer cells. Both LNCaP and PC-3 cells were plated in 100 mm dishes and treated with PFD for an additional two days. Cell lysates (50 µg) were separated by electrophoresis using a 12.5% SDS–polyacrylamide gel. After separation, proteins in the gel were transferred to a polyvinylidene difluoride membrane by electroblotting. Androgen receptor, prostate-specific antigen, phospho-Akt (Ser473), and total Akt protein levels were determined by Western blot analysis using specific antibodies. Equal loading of the samples was confirmed by measuring β-actin levels.