(B) RWPE-1 control cells and stably expressing AR wild type (AR WT) were either left untreated (C) or treated with 100ng/ml doxycycline (D) to induce ERG expression, in the absence (D) or presence (R/D) of 1nM R1881 for 24 hr. Western blot analysis shows expression levels of AR and ERG. (B) qRT-PCR of the luminal genes PSA (blue) and NKX3-1 (red) from RWPE-1 cells treated as in (A). Data represent normalized expression of PSA and NKX3-1 mRNA relative to the B2M transcript. Error bars represent + SEM of three replicates. (C) Western blot analysis of AR, ERG and GAPDH expression levels from RWPE-1 parental cells or cells stably expressing either AR wild type (WT), R711K, R727K, R753K, R761K, R775K, R780K, R787K, R789K, R787/789K, R832K, R841K, R847K, R855K, R856K, R855/856K or R872K AR mutant. (D) RWPE-1 cells stably expressing either AR R711K, R727K, R753K, R775K, R780K, R787K, R789K, R787/789K, R832K, R841K, R847K, R855K, R856K, R855/856K or R872K mutant were used for AR IP followed by western blot analysis for MMA, SDMA and total AR levels.