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), P_ERG6–OSH6 (FTY373), lag1Δ, P_ERG6–OSH6 lag1Δ (FTY527), vps13Δ, and P_ERG6–OSH6 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.