Figure 2.
The association of Smad3 with AR in a mammalian two-hybrid interaction system. (A) SW480.7 cells were cotransfected with 3 μg of Gal4-Smad3 encoding the full-length Smad3 fused to the Gal4-DBD and 4.5 μg of VP16-AR encoding the full-length AR fused to the activation domain of VP16. Interaction was estimated by determining the level of CAT activity from 3 μg of the reporter plasmid pG5-CAT in the presence of 10−8 M DHT. (B) DU145 cells were transfected with Gal4-Smad3 and VP16-AR expression vectors in the presence (+) or absence (−) of DHT and TGF-β. Each CAT activity is presented relative to the transactivation observed in the absence of DHT. All values represent the mean ± SD of four independent experiments.