Apg7p is present in the cytosol. A wild-type strain (YW5–1B) was cultured in MVD+Ura+Trp medium, transferred to nitrogen-starvation medium, and incubated for 30 min. Cells growing vegetatively were converted to spheroplasts in rich buffer (1% yeast extract, 2% polypeptone, 0.5% glucose, and 0.7 M sorbitol), whereas nitrogen-starved cells were converted to spheroplasts in nitrogen-starvation buffer (0.15% yeast nitrogen base without amino acids and ammonium sulfate, 2% glucose, and 0.9 M sorbitol). Cells were lysed and fractionated by centrifugation as described by Huang and Chiang (1997). LSP, 15,000 × g pellet; HSP, 100,000 × g pellet; HSS, 100,000 × g supernatant.