FIG. 9.

Inhibition of BAF57 or BRM attenuates AR-dependent proliferation. (A) LNCaP cells (10⁵) were transfected with 0.5 μg of pEGFP-H2B-GFP and 4.5 μg of parental vector, pBabe-BAF57ΔN, or dominant-negative AR (pSG5AR-Δ46-408). At 2 days posttransfection, cells were labeled with BrdU for 16 h and fixed. The percentage of transfected cells (H2B-GFP positive) incorporating BrdU was determined (left panel, representative image). The effect of dnAR or BAF57ΔN is shown as the percent inhibition of BrdU incorporation over vector control (right panel). (B) LNCaP cells were transfected with 0.5 μg of H2B-GFP and 4.5 μg of either pHTP-vector (lane 2) or pHTP-BRMi (lane 3). After transfection, cells were selected by using puromycin. Once cells were >90% H2B-GFP positive, they were lysed and subjected to immunoblot analyses for BRM and actin (loading control). SW13 cells (lane 1) are shown as a positive control for BRM deficiency. (C) LNCaP cells were transfected with 0.5 μg of H2B-GFP and 4.5 μg of parental vector, pHTP-BRMi scrambled, pHTP-BRMi, or pCMV-p16ink4a. The percentage of transfected cells (H2B-GFP positive) incorporating BrdU was determined. The effect of the BRM attenuation is shown as the percent inhibition of BrdU incorporation over vector control.