Autophagy-specific assays show COG4, TAX4, and SNX4 suppress the temperature-sensitive autophagy defect of the arl1Δ strain. (A) GFP-Atg8 degradation was increased at 37° in the arl1Δ strain when transformed with plasmids containing COG4, TAX4, and SNX4, as well as WT ARL1. All strains were cultured at 30° in the appropriate nonstarvation medium (containing either glucose or galactose) until log-phase, then all the strains were incubated at 37 or 30° for 30 min. The cells were then washed twice with SD-N (or SGal-N) medium, and cultured in SD-N (or SGal-N) for 3 hr either at 37 or at 30°. (B) COG4, TAX4, and SNX4 increased the Pho8Δ60 activities in arl1Δ (YSA003) at 37°. Error bars represent SD from three biological replicates. GFP, green fluorescent protein; SD-N, glucose nitrogen starvation medium; SGal-N, galactose nitrogen starvation medium; WT, wild-type.