BBR pro-degradative activity on ARpolyQ. (A) WB analysis on NSC34 cells transfected with AR.Q46 in absence or in presence of 10 nM testosterone. To inhibit protein synthesis the cells were pretreated with 20 μΜ cycloheximide (CHX) and then treated with 0.2 μΜ BBR or DMSO as vehicle control for 48 h. GAPDH was used as loading control. (B) The bar graph represents the mean optical density ± SD of AR: GAPDH (n = 3) (* p < 0.05, *** p < 0.001, two-way ANOVA, followed by Tukey’s test). (C,F) NSC34 cells transfected with AR.Q46 in absence or in presence of 10 nM testosterone and treated with 0.2 μΜ BBR and with 10 mM 3-MA (C,D) or 10 μΜ MG132 (E,F) to inhibit autophagic or proteasomal activity, respectively. (C,E) WB analyses were performed; GAPDH was used as loading control; the bar graphs represent the mean optical density ± SD of AR: GAPDH (n = 3). *** p < 0.001, two-way ANOVA, followed by Tukey’s test). (D,F) FRA was performed. The bar graphs represent the mean optical density of AR ± SD (n = 3). (* p < 0.05, ** p < 0.01, *** p < 0.001, two-way ANOVA, followed by Tukey’s test).