Panel A shows confocal microscopy images and fluorescence intensity graphs of mitochondrial protein colocalization. Panel B shows confocal microscopy images of mitochondrial apoptosis inducing factor expression across tissues. Panel C shows time lapse confocal microscopy images of green fluorescent protein after laser wounding. Panel D shows confocal microscopy images of mitochondrial apoptosis inducing factor accumulation on damaged mitochondria. Panel E shows time lapse confocal microscopy images of green fluorescent protein tagged mitochondrial apoptosis inducing factor. Panel F shows confocal microscopy images of mitochondrial membrane fluorescence using different emission channels. Panel G shows phylogenetic trees and graphs of predicted mitochondrial localization probabilities among homologs. Panel H shows confocal microscopy images and fluorescence intensity graphs of mitochondrial protein colocalization.
MAI-1 marks damaged mitochondria after laser wounding. (A) Left, representative confocal images of MAI-2::mKate2 and TOMM-20::GFP under basal conditions. Strains: Pcol-19-TOMM-20::GFP (zjuSi42);Pcol-19-MAI-2::mKate2 (zjuEx3210). Scale bar, 10 and 5 μm (zoom in). Right, quantification of fluorescence intensity along 15 μm lines across the images on the left. The white dashed lines indicate the analysis region. (B) Representative confocal images of Pmai-1::MAI-1::GFP expression in different tissues. Strains: Pmai-1-MAI-1::GFP (zjuSi602). Scale bar: 10 μm. The white arrowhead indicates the expression tissue. (C) Representative time-lapse confocal images of Pcol-19::GFP before and after laser wounding. Strains: Pcol-19::GFP (zjuSi60). Scale bar: 10 μm. The white dashed circle indicates the wound site. UW, unwounded; W, wounded. (D) Representative confocal images of MAI-1::GFP; mito::mKate2 before and after laser wounding. Strains: Pcol-19-mito::mKate2 (zjuSi47); Pcol-19-MAI-1::GFP (zjuEx2864). Scale bar: 10 and 5 μm (zoom in). The white dashed circle indicates the wound site. UW, unwounded; W, wounded. (E) Representative time-lapse confocal images of GFP::MAI-1 before and after laser wounding. Strain: Pmai-1-GFP::MAI-1 (zjuEx3038). Scale bar: 10 μm. UW, unwounded; W, wounded. (F) Representative confocal images of TOMM-20::mKate2 before and after laser wounding using 561 and 488 nm emission channel. Strains: Phyp-7-TOMM-20::mKate2 (zjuSi126). Scale bar, 10 μm. The white dashed circle indicates the wound site. UW, unwounded; W, wounded. (G) Phylogenetic analysis and predicted mitochondrial localization of ATP5IF1 homologs. Left, phylogenetic tree of ATP5IF1 homologs from multiple eukaryotic species, including nematodes, mammals, and early metazoans. Right, predicted probability of containing mitochondrial localization sequences. Proteins with a prediction probability of > 50% are considered to have a mitochondrial localization signal. (H) Left, representative confocal images of ATP5IF1::GFP and TOMM-20::mKate2 under basal conditions. Strains: Pcol-19-ATP5IF1::GFP (zjuEx3485);Phyp-7-TOMM-20::mKate2 (zjuSi126). Scale bar, 10 and 5 μm (zoom in). Right, quantification of fluorescence intensity along 15 μm lines across the images on the left. The white dashed lines indicate the analysis region.