FIG. 6.

BAF57 is recruited to the AR regulatory regions in the presence of ligand. LNCaP cells (5 × 10⁶) were cultured in steroid-free medium for 4 days and then stimulated with either 0.1% EtOH vehicle (lane 1) or 10 nM DHT (lane 2) for 60 min. After treatment the cells were formaldehyde cross-linked, and chromatin was recovered for use in ChIP assays. (A and B) Antibodies to acetylated histone H4 (AcH4), AR, BAF57h, and preimmune serum were used for ChIP analyses of the PSA enhancer region, as indicated. (C) A total of 10⁵ LB1-Luc cells (with integrated pARR2-Luc) were serum starved for 3 days and then treated with the indicated concentrations of DHT for 24 h. Reporter analysis was performed and demonstrated activation of the AR in a dose-dependent manner (left panel). For the ChIP analysis (right panel), LB1-Luc cells (5 × 10⁶) were cultured in steroid-free medium for 4 days and then stimulated with either 0.1% EtOH vehicle (lane 1) or 10 nM DHT (lane 2) for 60 min. After treatment, the cells were formaldehyde cross-linked, and chromatin was recovered for use in ChIP assays. Antibodies to AR and BAF57c were used for ChIP analyses of the ARR2 promoter region.