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. 2021 Nov 13;10(11):3161. doi: 10.3390/cells10113161

Figure 6.

Figure 6

The cyclin-dependent kinase Pho85 is critical for phosphate exhaustion-induced autophagy. (A) Schematic representation of the yeast phosphate signaling (PHO) pathway. (B–D) Survival during chronological aging, determined via flow cytometric quantification of PI staining, of WT, ∆rim15 and ∆pho81 cells (B), and of WT, ∆pho80 and ∆pho85 cells (C) grown on Ctrl or PR media; n = 4. Corresponding median lifespan is shown in (D). (E) Median lifespan of WT and ∆pho85 cells grown on Ctrl or nitrogen starvation (−N) medium; n = 4. (F) Confocal micrographs of WT and ∆pho85 cells harboring GFPAtg8 and Vph1RedStar (as vacuolar marker) at day 3 of aging on Ctrl or PR media. Scale bar represents 3 µm. (G,H) Representative immunoblot (G) and corresponding densitometric quantification (H) of protein extracts from WT and ∆pho85 cells harboring endogenously GFP-tagged Atg8 grown on Ctrl or PR medium. Blots were decorated with an antibody against GFP, and the ratio free GFP/GFPAtg8 was plotted; n = 8. For statistical analysis, Welch–ANOVA with Dunnett T3 post hoc test (D) or a two-way ANOVA (H) was used. * p < 0.05, ** p < 0.01 and *** p < 0.001.