Figure 6. Atg23 dimerization is critical for the role of Atg23 in the Cvt pathway and nonselective autophagy.
(A) atg23Δ cells exogenously expressing an empty vector, Atg23, or Atg23[LIL] under the endogenous promoter were grown overnight in nutrient-rich medium. Ape1 from cell lysates was inspected by western blot using anti-Ape1 antiserum.
(B) Quantification of (A).
(C) Empty vector, Atg23, or Atg23[LIL] driven by the endogenous promoter was expressed in atg23Δ cells also expressing GFP-Atg8 under the control of the CUP1 promoter. Cells were harvested in log phase after 0, 1, or 1.5 h of exposure to nitrogen-deficient medium. Processing of GFP-Atg8 to free GFP was assessed by western blot.
(D) Quantification of (C).
(E) Pho8Δ60 activity was assessed for atg23Δ Pho8Δ60 cells expressing empty vector, Atg23, or Atg23[LIL] under the control of the endogenous promoter. Quantification shows values as a percentage of wild-type activity after 4 h of nitrogen starvation.
(B, D, and E) Error bars represent SEM; n = 3; ***p < 0.001 by Tukey HSD test.