Figure 1.
Processing of GFP-Atg8 is blocked in the sec7 mutant. WT (SEY6210), atg1ts (WHY1 transformed with the plasmid expressing Atg1ts), and sec7ts (AFM69-1A) cells carrying the pCuGFPAtg8416 plasmid were grown at 24°C to an early log phase. Autophagy was then induced by addition of rapamycin, and cells were placed either at 24 or 37°C. Culture aliquots were collected at intervals of 1 h during a period of 4 h, and GFP-Atg8 cleavage was determined by Western blot analysis of the cell extracts. Bands were quantified using the Odyssey software, and the percentages of GFP-Atg8 (black) and GFP (gray) were plotted. Data for the sec7ts mutant graph represent the average of four experiments.