The Q-SNAREs Vam3, Vam7, and Vti1 act on the vacuole to promote autophagosome–vacuole fusion. (A) Fusion assay as in Fig. 2 D. Cytosol was prepared from the indicated deletions strains. The Sec18 inhibitor 5 mM N-ethylmaleimide (NEM) was added where indicated. (B) Fusion assay as in Fig. 2 D. Cytosol and autophagosomes were prepared from the indicated deletions strains. (C and D) Fusion assays as in Fig. 2 D. Cytosol, autophagosomes, and vacuoles were prepared from the indicated deletion or ts strains at 24°C. Fusion reactions were incubated at the restrictive temperature (30°C) for 2 h. All graphs show the mean from three independent experiments. Error bars are SD.