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. 2017 Nov 7;114(47):E10112–E10121. doi: 10.1073/pnas.1708367114

Fig. 1.

Fig. 1.

Atg20 is essential for the efficient initiation of bulk autophagy. (A) WT (WLY176) and atg20Δ (HPY063) cells expressing Pho8Δ60 were shifted from nutrient-rich conditions to SD-N medium for 4 h. (B) Autophagy as measured by the GFP-Atg8 processing assay in WT (SEY6210) and atg20Δ (D3Y009) cells. Cells transformed with the plasmid (pRS426) carrying a GFP-Atg8 construct were grown in rich selective medium and then starved for 1, 2, and 3 h. The free GFP:total GFP ratio was measured and normalized to that of WT after 3 h of starvation (100%). (C) Kinetics of prApe1 maturation in nutrient-rich medium and after the shift to SD-N medium for the indicated times. The vac8∆ atg20∆ (HPY079) strain was compared with the vac8Δ (CWY230) strain. The atg11Δ (SEY6210) strain served as a negative control, and Pgk1 served as a loading control. Quantitative evaluation of western blot data was done with three to four independent experiments carried out in nutrient-rich and nitrogen starvation conditions. Error bars represent the SD from three to four independent experiments. Statistical significance was tested using the unpaired two-tailed Student’s t test: **P < 0.01; ***P < 0.005.