Panel A shows super resolution microscopy images of ATP synthase and MAI-1 localization after mitochondrial damage. Panel B shows super resolution microscopy images of ATP synthase outer membrane proteins and MAI-1 colocalization. Panel C shows confocal microscopy images of reduced MAI-1 recruitment after ATP synthase gene silencing. Panel D shows quantification graph of MAI-1 fluorescence intensity changes following mitochondrial injury and RNA interference.
MAI-1 recruitment to damaged mitochondria requires ATP synthase. (A) Representative HIS-SIM single-plane images of ATP-3::GFP and MAI-1::mKate2 before and after ultrasonically induced damage. ROI, 1024 × 1024 pixels. Strains: ATP-3::GFP (zju133); Pcol-19-MAI-1::mKate2 (zjuSi594). Scale bar, 5 and 1 μm (zoom in). (B) Representative HIS-SIM single-plane images of ATP-3::GFP, TOMM-20::mYonghong, and MAI-1::BFP before and after ultrasonically induced damage. The white arrowhead indicates the membrane damage. ROI, 1024 × 1024 pixels. Strains: ATP-3::GFP (zju133); Pcol-19-TOMM-20::mYonghong (zjuSi577); Pcol-19-MAI-1::BFP (zjuSi539). Scale bar, 5 and 1 μm (zoom in). (C) Representative confocal images of MAI-1::GFP under L4440 (empty vector), atp-1, and atp-3 RNAi treatment 1 and 5 min after laser wounding. Strains: Pcol-19-MAI-1::GFP (zjuSi510). The white dashed circle indicates the wound site. UW, unwounded; W, wounded. Scale bar, 10 μm (zoom in). (D) Quantitation analysis of MAI-1::GFP intensity change at the injury site. n represents the number of wounds. Statistical analysis was performed using one-way ANOVA. Error bars represent the mean value ± SD (n = 50, 57, 54, 52, 56, 87, 59, 58, and 52 from left to right); ***, P < 0.001, compared with L4440.