Androgens induce miR-101 expression and miR-101 targets Ezh2. DHT-induced (25 nM, 24 hours) expression of miR-101 in presence or absence of flutamide (100 nM) in (a) mouse primary GC (mGC) culture, (b) KGN cell line, and (C) GCs isolated from mice injected with 10 mg DHT for 3 days. The average Ct values for miR-101 in media, DHT, and DHT plus flutamide–treated samples were 26.8/24.7/26.6 (mouse primary GCs), 25.6/23.6/26.3 (KGN cells), and 25.8/24.1 (DHT injections), respectively. (d–f) Effect of downregulation of miR-101 by anti-miR-101 inhibitor in primary mouse GC culture treated with DHT (25 nM, 48 hours) on (d) miR-101, (e) Ezh2 mRNA, and (f) Ezh2 protein levels. The average Ct values for miR-101 in media and anti-miR101–treated samples were 35.2 and 36.6, respectively. Data are displayed as mean ± standard error of the mean (n = 3 experiments) and normalized to U6 levels. *P ≤ 0.05.