Hydroxyflutamide (OH-Fl) reverses mibolerone’s effects on ER beta expression. (A) Real-Time RT-PCR for analyzing ER beta mRNA levels in cells treated as indicated. Each sample was normalized to GAPDH RNA content. Data represent the mean ± S.D. of values from three separate RNA samples expressed as the percentage of control assumed to be 100%. (B) Bottom panel, Western blot analysis of ER beta expression in cells treated with vehicle (−) or Mib 10 nM in the presence or absence of OH-Fl 1 μM for 48 hours. GAPDH was used as loading control. Upper panel, the histograms represent the mean ± S.D. of three independent experiments in which band intensities were evaluated in terms of optical density arbitrary units and expressed as the percentage of the control assumed to be 100%. *, P <0.01 compared to vehicle treated-cells. **, P <0.01 compared to Mib treated cells. ER, estrogen receptor.