Fig. 4

Mn does not affect mitochondria at low concentrations in N27 dopaminergic neuronal cells. (A,B) Mitochondrial respiration was assessed by measuring OCR using the XF^e96 Extracellular Flux Analyzer. At 62.5 and 125 µM, Mn did not impair basal respiration (P = 0.9859, P = 0.9997 respectively) or spare respiratory capacity (P = 0.3693, P = 0.8727 respectively) Data represent mean ± SEM, n = 3 independent experiments (C) N27 neuronal cells were treated with 125 µM or 250 µM of MnCl2 or vehicle control (RPMI media) for 24 h. Representative confocal images of mitochondrial morphology were captured after TOM20 immunostaining (upper panels), then skeletonized (lower panels) for subsequent analysis. Scale bar 20 μm. (D). Various parameters of mitochondrial morphology were quantified using Fiji MiNA plugin. No significant changes in branch length (P = 0.9082) or the number of branches per network (P = 0.9806) was detectable in 125 µM Mn treatment. Data represents mean ± SEM (n = 3 independent experiments with 30–40 cells per experiment), analyzed by Kruskal-Wallis ANOVA test