We also visualized the transport of GFP-Atg8 by fluorescence microscopy (Figure 2). Under nonstarvation conditions, GFP-Atg8 aggregated as single dots in the cells, denoting the PAS (Suzuki et al. 2001) (Figure 2A). When cells were treated with starvation medium at 30°, all of the strains had a vacuolar diffuse green phenotype (Yang and Rosenwald 2016), indicating the normal processing of GFP-Atg8. Under starvation conditions at 37° (Figure 2C), cells overexpressing COG4, SNX4, and TAX4 as well as ARL1 had the green diffuse phenotype, consistent with the conclusion from Figure 1 that these four genes, when overexpressed, are able to suppress the autophagy defect in the arl1Δ strain. On the other hand, cells overexpressing IVY1, PEP3, SLT2, and ATG5 exhibited multiple green dots, suggesting defective autophagy (Figure 2C). After counting the cells with diffuse green phenotype (normal autophagy) at both 30 and 37°, we found that the arl1Δ strains that overexpressed COG4, SNX4, and TAX4 had a significantly higher percentage of cells with the diffuse green phenotype at 37°, while cells overexpressing IVY1, PEP3, SLT2, and ATG5 had a dramatically decreased percentage of cells with this phenotype at high temperature compared with 30°. In conclusion, we identified COG4, SNX4, and TAX4 as high copy suppressors for the high temperature autophagy defect of the arl1Δ strain.