Fig. 7: Depletion of MTMR2 and Rubicon promotes autophagic clearance of damaged mitochondria.
From: Mitochondrial damage triggers the concerted degradation of negative regulators of neuronal autophagy
A Schematic depicting the steps of the mitophagy assay using DMP Red and Lysotracker to label acidified mitochondria and lysosomes, respectively. B, C Depletion of MTMR2 and Rubicon increases the number of mitochondria engulfed within autophagosomes. B Representative max projections of neurons nucleofected with Ctrl or Mtmr2 and Rubicon shRNA and assayed for mitophagy using the protocol represented in (A). Outline of the neuronal soma for quantification is indicated using dashed lines in cyan. Yellow boxes indicate inset regions. C Number of mitophagolysosomes per soma (marked by colocalizing DMP Red and Lysotracker punctae) of neurons nucleofected with Ctrl or Mtmr2 and Rubicon shRNA and assayed for mitophagy using the protocol represented in A (N = 4 experiments, two-tailed unpaired t-test). D–F Depletion of Rubicon is sufficient to significantly enhance mitophagic flux. D Representative max projections of neurons nucleofected with Ctrl, Mtmr2 or Rubicon shRNA and assayed for mitophagy using the protocol represented in (A). Outline of the neuronal soma for quantification is indicated using dashed lines in cyan. Yellow boxes indicate inset regions. E Number of mitophagolysosomes per soma (marked by colocalizing DMP Red and Lysotracker punctae) of neurons nucleofected with Ctrl, Mtmr2 or Rubicon shRNA and assayed for mitophagy using the protocol represented in (A). F Fold change in mitophagy flux per soma of neurons nucleofected with Ctrl, Mtmr2 or Rubicon shRNA and assayed for mitophagy using the protocol represented in (A) (D–F: N = 3 experiments, Kruskal–Wallis test). All panels: Error bars indicate SEM, scale bars = 5 μm. Source data are provided as a Source Data file.
