Fig. 5

Miro nanodomains associate with MICOS clusters. a–c Dual color dSTORM imaging of ^GFPMiro2-transfected HeLa cells. ^GFPMiro2 nanometer-sized domains are shown (anti-GFP : green) together with endogenous Mic60/Mitofilin (a) a positive control for Miro2 (b; anti-Miro2 antibody) and negative control for Tom20 (c), (scale bar: 0.2 μm). d Cross-correlation analysis between Miro2 and Mic60; Miro2 and ^GFPMiro2 and Miro2 and Tom20 and e Mean Van Steensel’s cross-correlation coefficient between ^GFPMiro2 and the different immunostainings calculated from (d) and plotted (n = cells; in which 5 ^GFPMiro2-Miro2, 9 Miro2-Mic60/Mitofilin, and 6 Miro2-Tom20 cells from three independent measurements were used; One-way ANOVA, Bonferroni post hoc). f Endogenous immunoprecipitation experiment in WT and DKO cells using antibodies against the core-forming MICOS components (Mic19/CHCHD3, Mic60/Mitofilin and Sam50). The main interactions are not to be critically affected. g, h Proximity ligation assay (PLA) and quantification in WT and DKO cells between Sam50 and Mic60/Mitofilin (g) and between Mic60/Mitofilin and Mic19/CHCHD3 (h) show a mild decrease in the association between these components (n = cells; in which 34 cells were used per condition from three independent experiments; Student’s t test with Welch’s correction), (scale bar: 10 μm). Error bars represent ± SEM. Significance: *p < 0.05; ***p < 0.001