Figure 6.

Atg8 does not control the frequency of autophagosome production. (A) GFP-Atg8 was expressed alone (strain GA8) or with additional Atg8 (strain GA8+A8). Samples were prepared as in Figure 5A. Protein extracts were analyzed by immunoblotting with anti-GFP antibody or anti-Atg8 antiserum. (B) Frequencies of autophagosome production in strains GA8 and GA8+A8 are comparable; the atg27Δ strain showed a significantly lower frequency. Yeast cells were starved for 1 h. The microscopy observation was performed as in Figure 3. The numbers of GFP-Atg8 peaks during 15 min were recorded. Error bar, SEM; n > 100. (C) Autophagy levels are different in strains GA8 and GA8+A8. The Pho8Δ60 assay was performed as in Figure 1. Error bar, SEM from three independent repeats.