Panel A shows confocal microscopy images of MAI-1 marked mitochondria undergoing gradual degradation after laser injury. Panel B shows confocal microscopy images of persistent MAI-1 accumulation in autophagy related mutant backgrounds. Panel C shows quantification graph of accumulated MAI-1 fluorescence in wild type and autophagy mutants. Panel D shows confocal microscopy images of MAI-1 and LGG-1 fusion during mitochondrial autophagy events.
MAI-1–marked mitochondria undergo autophagy-dependent degradation. (A) Representative confocal images of MAI-1::GFP and TOMM-20::mKate2 before and at different time points after laser wounding. Strains: Phyp-7-TOMM-20::mKate2(zjuSi126); Pcol-19-MAI-1::GFP (zjuSi510). Scale bar, 10 μm. (B) Representative confocal image of MAI-1::GFP in the epidermal hyp7 after wounding 5 min, 2 h, and 24 h in different genetic backgrounds. Mitophagy related mutants (pdr-1, lgg-1, and atg-3) were used. Strains: pdr-1(gk448), lgg-1(tm3489), atg-3(bp412), and Pcol-19-MAI-1::GFP (zjuSi510). Scale bar, 10 μm. (C) Quantitation of accumulated MAI-1::GFP in the wounding site in pdr-1(gk448), lgg-1(tm3489), and atg-3(bp412) after wounding 5 min, 2 h, and 24 h, shown in B. n represents the number of wounds. Statistical analysis was performed using one-way ANOVA. Error bars represent the mean value ± SD (n = 72, 67, 91, 78, 65, 36, 32, 92, 44, 64, 41, and 50 from left to right); ***, P < 0.001, compared with WT. (D) Representative confocal images of MAI-1::BFP and GFP::LGG-1 at different time points after wounding. The MAI-1 signal is shown in green, while the LGG-1 signal is shown in magenta for better visualization. The white dashed circle indicates the wound site. The white arrow indicates a fusion event. Strains: Pcol-19-GFP::LGG-1(zjuSi288); Pcol-19-MAI-1::BFP(zjuSi539). Scale bar, 5 μm. (A–D) UW, unwounded; W, wounded.