Back to article: Up-regulation of Osh6 boosts an anti-aging membrane trafficking pathway toward vacuoles

FIGURE 6: Up-regulation of OSH6 complements defects of lag1Δ. (A) Comparison of vacuole morphology of lag1Δ with the indicated plasmids listed on the left. Vector: YEp24, OSH4ox: YEp24-OSH4; OSH6ox: YEp24-OSH6. Log phase cells were labeled with FM4-64 for one hour and chased for three hours at 30°C and then photographed. (B) Quantitation of cells with different categories of vacuoles. Sample sizes are 98 for lag1Δ(vector), 105 for lag1Δ(OSH4ox) and 102 for lag1Δ (OSH6ox). A one-way ANOVA shows that the fraction with one vacuolar vesicle/cell of lag1Δ (OSH6ox) is significantly higher than that of lag1Δ (vector) (p< 0.0001). (C) Recovery of cell growth after caffeine and high temperature arrest. Overnight cultures of wild type (BY4742), PERG6OSH6 (FTY373), lag1Δ, PERG6OSH6 lag1Δ (FTY527), vps13Δ, and PERG6OSH6 vps13Δ (FTY534) were serially diluted by 10-fold (starting at 0.1 OD600/ml from the left). Five µl of serial diluted cells of the indicated strains were spotted on YEPD (left) and YEPD + 0.2% caffeine plate (middle) and incubated at 37°C for two days. Then the caffeine plate was incubated at room temperature (24°C) for five days.

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