A) Autophagy activity was measured by the Pho8Δ60 assay in WT, pat1Δ, ski2Δ, sk3Δ pat1Δ ski2Δ, and pat1Δ ski3Δ; strains under growing conditions (+N) and following 3 h of nitrogen starvation (−N). Error bars indicate the standard deviation of 6 independent experiments. ANOVA, **P <0.01 and *** P <0.001. B) Autophagy was measured by GFP-Atg8 processing in WT, pat1Δ, ski3Δ and pat1Δ ski3Δ strains under growing conditions and after 2 h of nitrogen starvation; a representative image is shown. C) Atg1, Atg2-PA, Atg7-PA and Atg9 protein levels were measured by western blot in WT, pat1Δ, ski3Δ and pat1Δ ski3Δ strains under growing conditions and after 1 or 2 h of nitrogen starvation; representative images are shown. D)
ATG1, ATG2, ATG7 and ATG9 mRNA levels were determined in WT, pat1Δ, ski3Δ and pat1Δ ski3Δ strains under growing conditions and after 1 h of nitrogen starvation by RT-qPCR. Error bars indicate the standard deviation of 3 independent experiments. ANOVA, ** P<0.01 and *** P <0.001. See also Figure S2.