Fig. 7.

Phosphorylation of Atg29 is required for binding to Atg11. (A) Plasmids encoding Atg29, Atg29[3SD], Atg29[3SE], or Atg29[23STA] were coexpressed with HA-Atg11 under the control of the CUP1 promoter (pCuHA-Atg11) in MKO ATG17-GFP (HCY107) cells. Cells were cultured in SMD and shifted to SD-N for 2 h, and cell samples were observed by fluorescence microscopy. The images are representative pictures from single Z-sections. DIC, differential interference contrast. (Scale bar, 2 μm.) (B) Quantification of Atg17-GFP dots. Twelve Z-section images were projected and the percentage of cells containing an Atg17-GFP puncta was determined. The SD was calculated from three independent experiments. (C) A plasmid encoding Atg29 or Atg29[23STA] was coexpressed with pCuHA-Atg11 in MKO ATG17-GFP (HCY107) cells. Cells were cultured in SMD, and cell lysates were prepared and incubated with IgG-Sepharose for affinity isolation.