Fig. 3: WIMF is dependent on mitochondrial Rho GTPase MIRO-1.

a Diagram of a genetic screen for outer mitochondrial membrane (OMM) proteins involved in WIMF. The candidate genes and mitochondrial morphological phenotype before and after wounding are listed in Supplementary Table 4. b Representative confocal images of epidermal mitochondria before and after needle wounding. N = 5 independent experiments. Arrows indicate the farthest extent of fragmented mitochondria from the wound site, which we used for the quantitation of distance of mitochondrial fragmentation. c Representative confocal images of epidermal mitochondria before and after Micropoint UV laser wounding (see also Supplementary Movie 6). Mitochondria were labeled by Pcol-19-mito::dendra2(juSi271) (b, c). Red mitochondria were photo-converted with a 405-nm laser. N = 3 independent experiments. Note WIMF is restricted to the injury site and does not spread to adjacent mitochondria in miro-1(tm1966) mutants. Scale bars (b, c), 10 μm. d Quantitation of the distance of fragmented mitochondria to the wound site after needle or Micropoint UV wounding in miro-1(tm1966) and rescued animals (WT-needle, n = 17; miro-1-needle, n = 24; WT-laser, n = 144; miro-1-laser, n = 99; GFP::miro-1( + )-laser, n = 68; miro-1; GFP::miro-1( + )-laser, n = 56 animals). Typically, we generated 20 arrows, as shown in panel b and averaged for each animal, see detail in methods. Bars indicate mean ± SEM. ****P < 0.0001, Two-tailed unpaired t-test. Source data are provided as a Source Data file. e Localization of GFP::MIRO-1 before and after needle wounding. Top, diagram of GFP::miro-1 knock-in strategy. Bottom, representative confocal image of Pcol-19-GFP::miro-1(zju21); Pcol-19-mito::mKate2(zjuSi47) (see also Supplementary Movie 7). GFP::MIRO-1 is localized around mito::mKate2 and was remained at the wounding site immediately after wounding. Scale bar, 5 μm and 2 μm (zoom-in). N = 2 independent experiments. f Fluorescence profiles from line scan 1 and 2 illustrate the presence of GFP::MIRO-1 on a section of the mitochondrial membrane before and after needle wounding. A.U. arbitrary units. Images representative of five animals. Note, the GFP signal surrounds the mitochondrial matrix mito::mKate2 signal.