Figure 1.

Arl1 and Ypt6 are involved in autophagy at the restrictive temperature of 37°C. (A) arl1Δ and ypt6Δ strains are sensitive to rapamycin at 37°C. All strains were streaked on YPD + DMSO (4.57 nM) or YPD + 5 ng/ml rapamycin in DMSO and cultured either at 30°C or 37°C. Plates were photographed after 3 d of growth. (B) GFP-Atg8 degradation was decreased in the arl1Δ strain at 37°C. WT (BY4743), atg1Δ, and arl1Δ strains were cultured at 30°C in nonstarvation medium until log-phase; then all the strains were incubated at 37°C or 30°C for 30 min. The cells were then washed twice with SD-N medium, and cultured in SD-N for 3 h either at 37°C or at 30°C. An aliquot of cells at 37°C were transferred back to 30°C and cultured in SD-N for an additional 3 h (denoted “R”). Whole cell lysates were subjected to immunoblot with anti-GFP antibody. (C) GFP-Atg8 degradation was decreased in the ypt6Δ strain at 37°C. The procedure was the same as Fig. 1B. (D) The arl1Δ (YSA003) and ypt6Δ (YSA004) strains show decreased Pho8Δ60 activity at 37°C. Error bars represent standard deviation from 3 biological replicates. The samples were done in 3 technical replicates for each biological replicate.