Fig. 6: Mitochondrial ROS is essential for neuroprotection.

a–c High-resolution respirometry measurements of the oxygen slope of isolated mitochondria treated for 10 min with CyPPA (10–50 µM). a measurements of state 2, basal respiration after addition of pyruvate, malate, glutamate. b complex I-linked respiration was determined after addition of ADP. c complex II-linked respiration after addition of succinate. Data are presented as mean ± SD, n = 3–5 independent experiments per condition. *p < 0.05, **p < 0.01. Statistical significance is assessed by one-way ANOVA Dunnet’s multiple comparisons. d Mitochondrial superoxides were measured using MitoSOX after CyPPA (10–50 µM, 24 h) treatment in the presence or absence of MnTBAP (20 µM). Data are presented as mean ± SD, n = 3, *p < 0.05, ***p < 0.001 versus untreated control, ^#p < 0.05, ^##p < 0.01 versus CyPPA only-treated cells with the corresponding concentration. e–h MTT assay of cells challenged with erastin, pre (8 h)- and co-treated with CyPPA (10 µM) in the presence or absence of MnTBAP (20 μM) in glucose (e) and galactose (f). The combination MnTBAP (20 μM) and DCA (10 mM) is depicted in g (glucose) and h (galactose). Data are presented as mean ± SD, n = 6, *p < 0.05, **p < 0.01, ***p < 0.001, ^##p < 0.01, ^###p < 0.001, ^$p < 0.05, ^$$p < 0.01, ^$$$p < 0.001, *compared to glucose or galactose control, ^#compared to erastin, ^$compared to erastin and CyPPA. All experiments were independently repeated at least three times.