Miro proteins coordinate microtubule‐ and actin‐dependent mitochondrial transport and distribution A, BMyo19 levels correlate with the quantity of Miro.

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Miro proteins coordinate microtubule‐ and actin‐dependent mitochondrial transport and distribution A, BMyo19 levels correlate with the quantity of Miro proteins in mouse fibroblasts. Western blot (A) and quantification (B) of Myo19 protein levels in all different genotypes. C, DMyo19 enrichment in the mitochondria is dependent on Miro proteins. (C) Western blot of endogenous Myo19 in the different cellular fractions from WT, Miro1KO, Miro2KO and MiroDKO MEFs. I: input; M: mitochondrial fraction; C: cytoplasmic fraction. (D) Enrichment of Myo19 on mitochondria calculated as mitochondrial signal/cytoplasmic signal and normalised to WT (n = 5 independent experiments; Kruskal–Wallis test with Dunn's correction). ERepresentative images of endogenous immunostained Myo19 and Mitotracker Orange in MEFs. FEnrichment of Myo19 in mitochondria (ratio of mitochondrial Myo19 signal to non‐mitochondrial signal) and comparison between WT (70 cells), Miro1KO (67 cells), Miro2KO (64 cells) and MiroDKO (54 cells) MEFs (n = 3 independent experiments; ANOVA‐NK). GRepresentative confocal images of cellular localisation of expressed GFPMyo19. HQuantified mitochondrial enrichment of GFPMyo19 signal (ratio of mitochondrial Myo19 signal to non‐mitochondrial signal) in the different MEF cell lines. Data come from the indicated number of cells from three different experiments (n = number of cells; ANOVA‐NK). I–LMiro proteins recruit Myo19 to the mitochondria. MiroDKO cells (I) and WT (K) expressing GFPMyo19 alone or together with the mitochondrial Miro2myc or the cytosolic Miro2ΔTMmyc (see Fig EV5C for the equivalent experiment with Miro1myc and Miro1ΔTMmyc). Quantification of mitochondrial enrichment of GFPMyo19 (ratio of mitochondrial signal to non‐mitochondrial signal) in the indicated conditions in MiroDKO (J) and WT (L). Data obtained from the indicated number of cells from three different experiments (n = number of cells; ANOVA‐NK). UT, untransfected. Data information: Error bars represent s.e.m. Statistical significance: *P < 0.05, **P < 0.01 and ***P < 0.001. Source data are available online for this figure. IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON PERMISSIONS@WILEY.COM OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER. EMBO J, Volume: 37, Issue: 3, Pages: 321-336, First published: 08 January 2018, DOI: (10.15252/embj.201696380)