Figure 10.

Localization of Atg1 and Atg9 in the atg17Δ mutant. (A) Wild-type (PSY143), atg17Δ (HCY032), atg13Δ (HCY040), and vac8Δ (HCY042) cells expressing Atg1-GFP were grown in YPD and shifted to SD-N for 2 h before microscopy. Atg1 showed both a punctate and diffuse cytosolic localization. The data for localization of Atg1-GFP in atg13Δ or vac8Δ were essentially the same as those shown for wild-type and atg17Δ. (B) Atg9 cycling is defective in cells lacking Atg17. Integrated Atg9-YFP was expressed in wild-type (FRY136), atg1Δ (FRY138), atg17Δ (KTY89), and atg1Δ atg17Δ (JLY4) cells. The cells were grown in YPD or appropriate selective media to mid-log phase and FM 4-64 was added to the culture medium for 15 min, after which the cells were pelleted and resuspended in YPD or SMD, and further incubated for 30 min, followed by imaging by fluorescence microscopy. In the merged panels Atg9-YFP is shown in green to facilitate visualization relative to the FM 4-64 dye. DIC, differential interference contrast.