FIGURE 3: Increased autophagic flux in [PSI⁺] strains.

(A) Autophagic flux was monitored in in the indicated strains expressing GFP-Atg8 during late exponential (day 1) and stationary phase growth (day 3). Increased free GFP is detected in the [PIN⁺][PSI⁺] strain indicative of autophagic flux. No free GFP is detected in the atg1 mutant strains which cannot initiate autophagy. Representative images are shown from triplicate experiments with quantification comparing the ratios of free GFP with GFP-Atg8 below. Significance is shown comparing the [PIN⁺][PSI⁺] versions of the wild-type and atg1 mutant strains with their corresponding [PIN⁺][psi^–] and cured [pin^–][psi^–] versions. Statistical analysis was performed by one-way ANOVA: *p < 0.05, **p < 0.01.

(B) Protein aggregates were isolated from the same strains as shown in panel A at day three of chronological growth and analyzed by SDS-PAGE and silver staining.