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. 2017 Jan 6;45(7):3738–3751. doi: 10.1093/nar/gkw1291

Figure 1.

Figure 1.

Androgen-stimulated AR Ser81 phosphorylation is associated with chromatin occupancy of BRD4 and P-TEFb. (A) LNCaP cells in steroid-depleted medium (RPMI with charcoal/dextran stripped serum, CDS medium) were pre-treated for 30 min with AR antagonist (bicalutamide: BIC, 10 μM; or enzalutamide: ENZ, 10 μM), CDK9 inhibitor (CDK9 inhibitor II: iII, 50 μM) or JQ1 (500 nM) as indicated, and then treated for 4 h with 10 nM DHT followed by lysis and immunoblotting. (B and C) LNCaP cells in CDS medium were treated for 4 h with DHT and BRD4 antagonist JQ1 (500 nM) as indicated, followed by immunoblotting (B) or RNA isolation and qRT-PCR analysis (C). (D and E) LNCaP cells in CDS medium were treated for 4 h with DHT and JQ1 as indicated, followed by ChIP-qPCR for occupancy of AR, pS81 (S81 phosphorylated AR), pS5 (S5 phosphorylated RNA Pol II), CDK9 and BRD4 on AR-regulated elements (AREs) in the KLK3/PSA (D) and KLK2 (E) genes.