Figure 5.

Enforced expression of Smad3 reverses the protective effect of DHT on TGF-β1-induced cell death. A, NRP-154+AR cells were infected with either control virus or AdMax-Flag-Smad3 (1:500) and treated in GM3 medium with TGF-β1 (10 ng/ml) in the absence or presence of three different doses of DHT. After 5 d of TGF-β1 treatment, medium was replaced with GM2.1 medium with/without DHT and further cultured for 1 wk. B, NRP-154 cells were transiently infected with Admax-AR (1:8000) along with either Admax-cont, Admax-Smad3 (1:500), or AdMax-TβRII and then treated with TGF-β1 (5 ng/ml for 72 h or 96 h) or without TGF-β in the absence/presence of DHT (10 nm). After TGF-β1 treatment, cells were washed and cultured in growth medium, GM2, for an additional 6 d. Cells were then washed/fixed and stained with crystal violet, after which stained colonies were photographed (A) or dye was eluted to measure cell viability by reading the optical density at 550 nm (B). Each bar represents the average of triplicate determinations ± se. cont, Control; wt, wild type.