Figure 6

Inhibition of NOX complexes reduces ROS production, increase the GSH level and ameliorates toxicity. A) Measurement of mitochondrial superoxide levels using MitoSox in cells induced to express ATXN7Q65-GFP for 0, 6, 9 or 12 days. Antimycin A treatment was used as a positive control. B) Effect of NOX complex inhibition on ATXN7Q65-GFP cells. ROS levels (top panel), GSH levels (middle panel) and cell viability (lower panel) was analyzed in FLQ65 cells not induced (+Dox) or induced (−Dox) to express ATXN7Q65-GFP for 9 days while growing in media with or without the NOX complex inhibitor apocynin (50 μM). C) Effect of NOX inhibition by apocynin on ATXN7 aggregation and ATXN7Q65-GFP expression in FLQ65 cells grown and treated as in A. Top panel; representative western blot and quantification of expression. Lower panel; representative dot blot and quantification of aggregation. D) Effect of NOX complex inhibition by gp91ds-TAT on ATXN7Q65-GFP cells. ROS levels (top panel) and cell viability (lower panel) was analyzed in FLQ65 cells not induced (+Dox) or induced (−Dox) to express ATXN7Q65-GFP for 9 days while growing in media with or without 10 μM of the inhibitor peptide. All quantifications are shown as means ± SEM from three independent experiments with triplicates. NS: not significant, * p <0.05, ** p <0.01 and *** p <0.001.