SAMM50-dependent basal piecemeal mitophagy requires p62 and hATG8 proteins. (A and B) Immunoblots of extracts of WT and p62 KO HeLa cells treated with BafA1 for 24 h (A) and quantified (B). Values are mean ± SD from three different experiments. **, P < 0.005; *, P < 0.01; one-way ANOVA. (C) Time-lapse (seconds) live-cell confocal imaging of HeLa cells stably expressing mCherry-p62 and MIC19-EGFP. Arrows show mitochondrial fragment colocalized with p62 and subsequently degraded (see Video 1). Scale bar, 5 µm. (D) Time-lapse (seconds) live-cell confocal imaging of HeLa cells stably coexpressing LAMP1-EGFP and MIC19-mCherry. Arrows indicate mitochondrial fragment engulfed by a lysosome (see Video 2). Scale bar, 2 µm. (E) Live-cell images of HeLa cells stably expressing MIC19-Keima excited at 458 nm in a neutral environment (mitochondria; green) and at 561 nm in an acidic environment (lysosome; red). Scale bars, 10 µm (main), 2 µm (inset).
Figure 5.

SAMM50-dependent basal piecemeal mitophagy requires p62 and hATG8 proteins. (A and B) Immunoblots of extracts of WT and p62 KO HeLa cells treated with BafA1 for 24 h (A) and quantified (B). Values are mean ± SD from three different experiments. **, P < 0.005; *, P < 0.01; one-way ANOVA. (C) Time-lapse (seconds) live-cell confocal imaging of HeLa cells stably expressing mCherry-p62 and MIC19-EGFP. Arrows show mitochondrial fragment colocalized with p62 and subsequently degraded (see Video 1). Scale bar, 5 µm. (D) Time-lapse (seconds) live-cell confocal imaging of HeLa cells stably coexpressing LAMP1-EGFP and MIC19-mCherry. Arrows indicate mitochondrial fragment engulfed by a lysosome (see Video 2). Scale bar, 2 µm. (E) Live-cell images of HeLa cells stably expressing MIC19-Keima excited at 458 nm in a neutral environment (mitochondria; green) and at 561 nm in an acidic environment (lysosome; red). Scale bars, 10 µm (main), 2 µm (inset).

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