Panel A shows confocal microscopy images of lysosomes approaching and degrading MAI-1 positive mitochondria after injury. Panel B shows time lapse confocal microscopy images of lysosome fusion with damaged MAI-1 labeled mitochondria. Panel C shows confocal microscopy images and fluorescence intensity graphs of lysosome colocalization with damaged mitochondria. Panel D shows time lapse confocal microscopy images of SCAV three mediated engulfment of damaged MAI-1 labeled mitochondria.
Lysosomes engulf MAI-1–marked mitochondria after injury. (A) Representative confocal images of MAI-1::GFP and NUC-1::mCherry at different time points after laser wounding. Lysosomes approached and degraded MAI-1–positive mitochondria. Strains: Pced-1-NUC-1::mCherry(qxIs257); Pcol-19-MAI-1::GFP(zjuSi510). Scale bar, 10 and 5 μm (zoom in). (B) Time-lapse confocal images of MAI-1::GFP and NUC-1::mCherry after precise single mitochondrial wounding. Lysosome fusion coincides with MAI-1 signal loss. The white dashed circle indicates the wound site. The white arrowhead indicates a fusion event. Strains: Pced-1-NUC-1::mCherry(qxIs257); Pcol-19-MAI-1::GFP(zjuSi510). Scale bar, 5 μm. (C) Left, representative confocal images of TOMM-20::mYonghong, NUC-1:: mNeonGreen, and MAI-1::BFP before and 2 h after laser-induced wounding. The white arrow indicates colocalized MAI-1–positive mitochondria and lysosome. Right, quantification of fluorescence intensity along 15 μm lines across the wound site. Strains: Pcol-19-NUC-1::mNeonGreen (zjuSi534); Pcol-19-MAI-1::BFP (zjuSi539); Pcol-19-TOMM-20::mYonghong (zjuSi577). Scale bar, 10 and 5 μm (zoom in). (D) Representative time-lapse confocal images of MAI-1::mKate2 and SCAV-3::GFP before and at different time points after laser wounding. The white arrowhead indicates an engulfing event. Strains: Pscav-3-SCAV-3::GFP (qxIs430); Pcol-19-MAI-1::mKate2(zjuSi594). Scale bar, 5 μm. (A–D) UW, unwounded; W, wounded.