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. 2023 Aug 28;42(19):e112814. doi: 10.15252/embj.2022112814

Figure 7. Ypt1 is an autophagy determinant controlled by TOR.

Figure 7

  • A–H
    (A, E) AID‐3×HA‐Ypt1 yeast strains co‐expressing GFP‐Atg8 and Vph1‐mCherry were transformed into empty vector, FLAG‐Ypt1, FLAG‐Ypt1 S174A , or FLAG‐Ypt1 S174D plasmids. These yeast strains were treated with 0.5 mM IAA for 2 h to degrade endogenous AID‐3×HA‐Ypt1 for 2 h, and then were subject to nitrogen starvation for 1 or 4 h. Images of cells were obtained using an inverted fluorescence microscope. Scale bar, 2 μm. (B, F) Cells from (A, E) were assessed for the translocation of GFP‐Atg8 into vacuoles. n = 300 cells were pooled from three independent experiments. Data are presented as mean ± SD. ***P < 0.001; **P < 0.01; ns, no significance; two‐tailed Student's t‐tests were used. (C, G) The autophagic activity of cells from (A, E) were analyzed by western blot for the cleavage of GFP‐Atg8. Pgk1 served as a loading control. (D, H) The cleavage of GFP‐Atg8 from (C, G) were quantified and presented as mean ± SD (n = 3). ***P < 0.001; **P < 0.01; NS, no significance; two‐tailed Student's t‐tests were used.

Source data are available online for this figure.