We performed the same set of autophagy assays as for the arl1Δ strain to test which of the seven genes from the screen were able to suppress the high temperature autophagy defect. From the GFP-Atg8 assay, we found that only IVY1 and ATG5, as well as the positive control YPT6, were able to suppress the autophagy defect of the ypt6Δ strain at 37°, as demonstrated by the appearance of free GFP (Figure 3A). This conclusion was further confirmed through Pho8Δ60 assay (Figure 3B); upon overexpression of IVY1, ATG5, or YPT6, the Pho8Δ60 activity in the ypt6Δ strain (YSA004) was increased compared with empty vector. Similarly, these results were confirmed by the GFP-Atg8 fluorescence phenotype (Figure 4, A–C), as we found that the ypt6Δ strain overexpressing IVY1 or ATG5 had an increased percentage of cells with the diffuse green phenotype at 37° (Figure 4D). In conclusion, we identified IVY1 and ATG5 as high copy suppressors for the high temperature autophagy defect of the ypt6Δ strain.