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. 2022 Dec 2;15:966209. doi: 10.3389/fnmol.2022.966209

Figure 1.

Figure 1

Miro1 R272Q does not change mitochondrial movement or Miro1 degradation upon CCCP-induced mitophagy. (A) Representative image of hDaNs stained with neuronal marker MAP2 and dopaminergic marker TH; nDiff = 3. Scale bar: 50 μM. (B) Miro1 protein levels in hDaN lysates. Representative blot and quantification of intensity of Miro1 bands relative to a-tubulin. nDiff = 8, data displayed as mean ± SD; paired t test (two-tailed). (C) Miro1 and Mitofusin protein levels in hDaNs upon induction of mitophagy using 10 μM CCCP for 0/2/4/6 (+ 10 μM MG132)/22 h. Representative blot and quantification of intensity of Miro1 and Mitofusin bands relative to a-tubulin. nDiff = 3, data displayed as mean ± SD; Miro1: Two-way ANOVA with Tukey’s multiple comparisons; Mitofusin: Friedman test with Dunn’s multiple comparisons. (D) Representative first frame of movie and derived kymograph for movement analysis of hDaNs stained with 100 nM MitoTracker green. Scale bar: 10 μM. (E) Kymograph analysis of mitochondrial movement to classify stationary/oscillating/anterograde/retrograde fractions and mean displacement of mitochondria. nDiff = 3 (2 datasets per differentiation), data displayed as mean ± SD; Fractions: Two-way ANOVA with Šídák’s multiple comparisons; Displacement: Friedman test with Dunn’s multiple comparisons. (F) Mean mitochondrial speed analyzed using TrackMate fiji plugin. nDiff = 3 (nProcesses = 60), data displayed as mean ± SD; Mann–Whitney test.