Figure 4. Ypt1‐Atg23 binding is required for the PAS localization of Atg9 vesicles.

• A
  Wild‐type yeast strains co‐expressing Atg17‐2×mCherry and Atg9‐2×GFP or AID‐3×HA‐Ypt1 yeast strains co‐expressing Atg17‐2×mCherry and Atg9‐2×GFP transformed with empty vector, FLAG‐Ypt1, or FLAG‐Ypt1^W62A plasmids were treated with IAA for 2 h to degrade endogenous AID‐3×HA‐Ypt1, and then subjected to SD‐N for 0 or 1 h. Images of cells were obtained using fluorescence microscopy. Scale bar, 2 μm.
• B
  Cells from (A) were quantified for the number of cells in which Atg9‐2 × GFP colocalized with Atg17‐2 × mCherry puncta. n = 300 cells were pooled from three independent experiments. Data are shown as mean ± SD. ***P < 0.001; ns, no significance; two‐tailed Student's t‐tests were used.
• C
  Cells from (A) were quantified for the number of Atg9‐2 × GFP. n = 300 cells were pooled from three independent experiments. Data are shown as mean ± SD. ns, no significance; two‐tailed Student's t‐tests were used.
• D–F
  AID‐3×HA‐Ypt1 cells co‐expressing Atg9‐TAP and Atg11‐2×GFP, Atg13‐2×GFP, or Atg17‐2×GFP transformed with empty vector, FLAG‐Ypt1, or FLAG‐Ypt1 ^W62A plasmids were grown to log phase, and then subjected to SD‐N for 1 h. Cell lysates were immunoprecipitated with anti‐GFP agarose beads and then analyzed by western blot using the indicated antibody.

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