Figure 2.
Combination of enzalutamide and 17-AAG led to decreased AR protein level and transcriptional activity. (A&B) LNCaP (A) and C4-2 (B) cells were treated as indicated for 24 hr, followed by IB against AR, PSA and CHIP. (C&D) 22RV1 (C) and MR49F (D) cells were treated as indicated for 24 hr, followed by IB against AR and HSP90. (E) C4-2 cells were treated as indicated for 24 hr, fractionated into cytoplasm and nuclear, followed by IB against AR and Plk1. (F) C4-2 cells were treated as indicated for 24 hr, and crosslinked with 1% formaldehyde. The chromatin fraction was separated and subjected to anti-histone H3 immunoprecipitation (IP), followed by IB against AR. (G) C4-2 cells were treated as indicated for 24 hr and subjected for CHIP analysis using AR antibody. Binding of AR to the promoter region of PSA, CAMKK2 and FKBP5 were measured using RT-PCR with specific primers targeting the promoter region of indicated genes.