Figure 3.

Activation of the Golgi Arf proteins by the GEF activity of Sec7, Gea1, and Gea2 is essential for autophagy. The erg6Δ, gea1^ts gea2Δ (APY022), arf1^ts arf2Δ (SSY146), ret2^ts (PC130), and ret3^ts (FLY89) cells carrying the pCuGFPAtg8416 or the pCuGFPAtg8414 plasmid were grown and processed as described in Figure 1, with the exception of erg6Δ, which was analyzed at 30°C. To inhibit all the Sec7 GEF activity, BFA was added to the erg6Δ mutant. Quantification of the GFP-Atg8 processing was done using an Odyssey system. Graphs represent an average of two experiments. Identical results were obtained when the same experiments were repeated, and autophagy was induced by nitrogen starvation in the SD-N medium (data not shown).