Figure 1.

Basal and nitrogen starvation-induced autophagy normally occur in mmi1Δ cells. (A) Exponentially growing WT, atg8Δ, and mmi1Δ cells expressing Pho8Δ60 (OD₆₀₀ ≈ 0.8) were shifted to nitrogen starvation medium (SD-N). Samples were taken in indicated time points, proteins extracted, and the specific Pho8Δ60 activity was determined. Results are means ± SD of three independent experiments performed in duplicates (n = 6). The statistical evaluation was performed by using two way analysis of variance (ANOVA). The threshold for significance was set as p ≤ 0.01. NS; not significant; (B) Western blot detection of GFP-Atg8 cleavage in the WT, atg1Δ, and mmi1Δ cells. Cells expressing GFP-Atg8 were grown until the logarithmic growth phase (OD₆₀₀ ≈ 0.8) and then shifted to the SD-N medium. Samples were taken at indicated time points and the cleavage of GFP-Atg8 was analyzed by Western blot detection with antibodies against GFP. Detection of Pgk1 was used as a loading control. Quantification of the blots is presented below. GFP ratio (free GFP/Pgk1) was calculated. Error bars reflect SD from the two independent experiments.