Figure 6.

Atg13-Vac8 binding is required for correct initiation complex localization and robust autophagy activity. (A) Autophagy activity was measured using the Pho8Δ60 assay in strains expressing either wild-type Atg13-GFP or truncated Atg13-GFP[1–659], the vac8∆ strain expressing truncated Atg13-GFP[1–659], or vac8∆ or atg13∆ strains under nutrient-rich conditions (+N) and after 3 h of nitrogen starvation (-N 3 h). Error bars indicate the standard deviation of 5 independent experiments. ANOVA, *P < 0.05; **P < 0.01; ***P < 0.001. ns, no significance. (B) Atg13-GFP and truncated Atg13-GFP[1–659] association with the vacuole was determined using fluorescence microscopy. The vacuolar membrane was stained using the dye FM 4–64. (C) Quantification of panel (B). The percentage of cells in which Atg13-GFP puncta associated with the vacuole is presented. Student’s t-test, ***P < 0.001