Figure 5.

Targeting Vac8 to peroxisomes recruits Atg13 and increases pexophagy. (A) Atg13-GFP and either wild-type Vac8-RFP or peroxisome-targeted Vac8-RFP colocalization was determined using fluorescence microscopy. The vacuolar lumen was stained using CMAC. (B) Pexophagy activity was measured with the Pex14-GFP processing assay in Vac8-RFP WT and Vac8-RFP Pex strains, after 0, 2 and 3 h of nitrogen starvation (-N). A representative image is shown. Duplicate sets of samples are shown for each strain. (C) Quantification of panel (B). The ratio of free GFP to Pex14-GFP after 3 h of nitrogen starvation (-N) is presented. Error bars indicate the standard deviation of 5 independent experiments. Student’s t-test, ***P < 0.001. (D) Pexophagy activity was measured using the Pex14-GFP processing assay in WT and atg36∆ cells expressing either Vac8-RFP WT or Vac8-RFP Pex strains, after 0 and 4 h of nitrogen starvation (-N). A representative image is shown. (E) Quantification of panel (D). The ratio of free GFP to Pex14-GFP after 0 and 4 h of nitrogen starvation (-N) is presented. Error bars indicate the standard deviation of 4 independent experiments. ANOVA, *P < 0.05; ***P < 0.001