Fig. 6.

Knockdown of SRD5A2 results in a significant decrease in androgen receptor target gene expression in cells with active NF-κB but not in cells overexpressing AR-V7. NHPrE1-EE (constitutively active NF-κB—A) and NHPrE-V7 (expressing AR-V7—B) cell lines transfected with a control siRNA (control) or with siRNA corresponding to SRD5A2 (si-SRD5A2) were incubated in the presence of vehicle or testosterone (T). AR target gene expression was examined 12 hr later. Expression is relative to empty vector control cells that do not express androgen receptor. (A) qPCR of NHPrE1-EE for AR target genes PSA (right) and TMPRSS2 (left) resulted in a significant increase in mRNA expression in the presence of androgens when compared to vehicle (P < 0.001). SRD5A2 knock down cells had a significant decrease in PSA and TMPRSS2 expression even in the presence of androgens when compared to EE (P < 0.01) and lost the ability to respond to androgens when compared SRD5A2 knock down vehicle treated. (B) qPCR inNHPrE1-V7 cell lines for AR target genes PSA and TMPRSS2 resulted in no change in mRNA expression in the presence of testosterone. Bars are presented as standard deviation, P-value: two-way ANOVA. All experiments were performed three times with triplicate repetitions (n = 9).