Figure 2.

AR S81 phosphorylation and p300 recruitment coordinate a positive-feedback loop to sustain AR-mediated transactivation. (A) LNCaP cells in CDS medium were treated for 4 h with DHT (10 nM) and CDK9 antagonist (iII: CDK9 inhibitor II, 50μM), followed by ChIP-qPCR for p300 binding to the AREs in the KLK3/PSA and KLK2 genes. (B) LNCaP cells in CDS medium were treated with DHT and p300 antagonist (C646, 10 μM), followed by ChIP-qPCR for binding to AR-regulated PSA and KLK2 enhancers. H3K27Ac: Lysine 27 acetylated histone H3. (C and D) LNCaP cells in CDS medium were treated with DHT and C646, followed by RNA isolation for qRT-PCR (C) or immunoblotting (D).