Fig. 1

Endpoint methods such as serial dilution and colony formation assays fail to detect a nitrogen-starvation sensitivity phenotype for atg27Δ mutants. a WT, atg27Δ, and atg1Δ yeast strains were serially diluted and spotted on YEPD medium after 0 or 14 days of SD-N treatment (nitrogen starvation). The WT and atg27Δ strains were indistinguishable from one another despite published biochemical evidence that atg27Δ strains have autophagic marker processing and autophagosome formation defects. As expected, atg1Δ displayed a strong starvation sensitive phenotype that was easily noticeable when compared to WT. b Colony formation units were quantified after SD-N treatment of WT, atg27Δ, and atg1Δ yeast strains for the indicated period of time. The percentage of colonies produced by the same OD₆₀₀ and volume of culture at day 0, 7, or 14 post-starvation (% viability) was calculated and plotted as a function of nitrogen starvation time (days). No statistically significant difference was observed between the starvation sensitivity of WT and atg27Δ. All experiments were carried out at least three times, with at least three technical replicates for each experiment. While this plot was generated using a set of three technical replicates, all experiments displayed the same trends. Error bars indicate standard deviations, and p-values less than 0.05 are flagged with one asterisk (*)