Figure 3.

Arl1 and Ypt6 are required for Atg8 transport to the vacuole at 37°C. Atg8 is mislocalized in arl1Δ and ypt6Δ strains at 37°C. Cells were grown, then starved for nitrogen as described. FM 4–64 was used to stain the vacuolar membrane. Experiments were repeated 3 times and the results shown are from a single experiment. (A) Fluorescence images for WT, atg1Δ, arl1Δ and ypt6Δ in nonstarvation conditions. (B) Fluorescence images for WT, atg1Δ, arl1Δ and ypt6Δ in starvation conditions at 30°C. (C) Fluorescence images for WT, atg1Δ, arl1Δ and ypt6Δ in starvation conditions at 37°C. (D) The percentage of cells with Atg8 dots in 3 categories: 0, ≤ 2 and multiple dots (> 2 dots per cell) was quantified. At least 150 cells were counted for each strain. Error bars represent standard deviation from 3 biological replicates. Scale bar: 3 µm.