Figure 6.

Effect of retromer on autophagic activity. The level of prApe1 maturation was determined in immunoblots (A) and the percentage of mApe1 was calculated (B). Autophagic breakdown of GFP-Atg8 (C, D) as well as a Pgk1-GFP assay to determine nonselective autophagy (E,F) was analyzed in immunoblots and the amount of rather proteolysis stable GFP was quantified. Cells were nitrogen starved in SD-N and harvested for analysis. The prApeI maturation rate was determined by dividing the amount of mature ApeI to the total amount of Ape1, while the ratio of free GFP to total GFP was calculated for both the GFP-Atg8 and Pgk1-GFP assay. At least three independent experiments were quantified. Statistical relevance was determined using an unpaired two-tailed t-test, error bars indicate SEM, asterisks indicate p-values.