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. Author manuscript; available in PMC: 2012 Jul 22.
Published in final edited form as: Cell. 2011 Jul 22;146(2):290–302. doi: 10.1016/j.cell.2011.06.022

Fig. 7. Autophagy Is Impaired in the Absence of Sec1, Sec18, and the R/v-SNAREs Sec22 and Ykt6.

Fig. 7

(A, C and F) The indicated mutant and the corresponding wild-type (BY4742) strains were analyzed with the GFP-Atg8 processing assay in rich or nitrogen starvation conditions. R, recovery.

(B, D and G) vam3ts (UNY162), atg1Δ (TYY164), sec18-1, sec22-1 and ykt6ts cells expressing GFP-Atg8 were pre-incubated at 34°C for 30 min, shifted to starvation conditions for 1 h at the NPT and examined for sensitivity to proteinase K (PK) with or without 0.2% Triton X-100 (TX) as described in the Supplement.

(E and H) Cells expressing GST-Sec9 under the control of the GAL1 promoter (UNY172) were transformed with the indicated plasmids; CUP1 promoter-driven PA-Sec22, PA-Ykt6 or PA-Gos1, and GFP-Sso1, or CUP1 promoter-driven PA alone and GFP-Sso1. Cells were grown in SMG medium medium to OD600 = 1.0 and shifted to SG-N for 1 h. Spheroplasts prepared from these cells were subjected to DSP cross linking as described in the Supplement. Cell lysates were prepared and subjected to affinity isolation with IgG sepharose. The asterisk indicates a non-specific band. See also Figure S7.