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. 2021 Aug 9;31(1):111–121. doi: 10.1093/hmg/ddab225

Figure 1.

Figure 1

Interaction of Hsv2 with Atg2. (A) Two-hybrid assays. GAD-Hsv2 fusion was tested for two-hybrid interaction with GBD-Atg2 and GBD-Hsv2. Strains were Y187 or OVY546 (Δatg18). Values for ß-galactosidase activity are averages for eight transformants. The mean values are shown with standard deviation (SD). Values for two-hybrid control tests were <0.05 U. (B) Atg2-3mCherry complements the autophagic defect of a Δatg2 mutant. Y00000 (WT) and OVY384 (Δatg2) were cotransformed with pGFP-Atg8 and either pAtg2-3mCherry (+) or an empty vector (−). Cells were grown to mid-log phase and starved 4 h in SD-N medium. Protein extracts were immunoblotted with anti-GFP Ab to detect GFP-Atg8 and free GFP. (C) Colocalization of Hsv2 and Atg2. OVY545 expressing Hsv2-GFP and Atg2-3mCherry was grown to mid-log phase (− Rap) or treated with rapamycin for 90 min (+ Rap) and examined by fluorescence and differential interference contrast (DIC) microscopy, Scale bar: 1 μm. The percentage of Atg2 puncta colocalized with Hsv2 is shown on the right. The graph represents the mean of three experiments ± SD. *P < 0.05.