FIG. 2.

BAF57 binds the AR independent of ligand. (A) Immobilized GST or GST-BAF57 was incubated with [³⁵S]methionine-labeled AR in the presence of 0.1% EtOH vehicle (lanes 2 and 3) or 10 nM DHT (lanes 4 and 5) for 3 h at 4°C, washed, and denatured. Input and bound protein was subjected to SDS-PAGE, and ³⁵S-labeled AR was detected by autoradiography. (B) Clarified protein lysates from LNCaP cells was utilized for coimmunoprecipitation with antibodies to AR (lane 1), BAF57c (specific to C terminus of BAF57) (71) (lane 2), or preimmune serum (lane 3). The resultant immunoprecipitates were subjected to SDS-PAGE, and the presence of AR was detected by immunoblotting.