Figure 3.

In vivo and in vitro interaction between Smads and AR. (A and B) Coimmunoprecipitation of AR and Smad3. (A) PC-3 cells that overexpressed Flag-Smad3 and AR. (B) PC-3 and PC-3(AR)2 cells were treated with or without DHT. Cell extracts were prepared and immunoprecipitations were performed with the use of anti-FLAG antibody or anti-Smad3 antibody, followed by immunoblotting with antibody to AR. (C) The wtAR and different AR deletion mutants used in the GST pull-down assay are shown schematically. (D) Interaction domains of AR for Smad3. A series of ³⁵S-labeled mtARs incubated with GST-Smad3 or GST alone in the presence (+) or absence (−) of 1 μM DHT were tested for interaction in the GST pull-down assay.