Fig. 2.

Vps13 is required for cortical ER-phagy but not nuclear ER-phagy. (A) The translocation of Rtn1-GFP to the vacuole is disrupted in the vps13Δ mutant. (A, Left) Fluorescence images of yeast cells expressing Rtn1-GFP. Cells were treated with rapamycin for 16 h. Vacuoles were stained with FM4-64. (A, Right) The percentage of cells with GFP in the vacuole was quantified. Error bars represent SEM; N = 3 independent experiments. **P < 0.01, ***P < 0.001, Student’s t test. (B) The degradation of Rtn1-GFP in the vacuole is disrupted in the vps13Δ mutant. (B, Left) Cells expressing Rtn1-GFP were treated with rapamycin for 0 or 16 h. Cell lysates were immunoblotted using an anti-GFP antibody. Adh1 was used as a loading control. (B, Right) The percentage of free GFP divided by total GFP was quantified. The data were normalized to the WT control. Error bars represent SEM; N = 3 independent experiments. **P < 0.01, ****P < 0.0001, Student’s t test. (C, Left) Western blot analysis of the cleavage of Hmg1-GFP to GFP after rapamycin treatment for 0 or 16 h. Adh1 was used as a loading control. (C, Right) The percentage of free GFP divided by total GFP was quantified. The data were normalized to the WT control. Error bars represent SEM; N = 3 independent experiments. NS, nonsignificant (P ≥ 0.05), ***P < 0.001, Student’s t test. (Scale bar, 2 μm.)