Fig. 3

EVT involves a conjugation-independent function of Atg8. a, b The degradation of GFP-Pho8 is independent of most ATG genes except for ATG8. Cells were treated as Fig. 1a. Data are presented as in Fig. 1a and b, respectively. a atg8Δ cells accumulated vesicle-like structures within vacuoles. b atg8Δ cells contained less free GFP and accumulated a truncated form of Pho8. See Fig. S2A for quantification. c Cytosol is sequestered in the intravacuolar structures in atg8Δ cells. atg8Δ cells co-expression GFP-Pho8 and DsRed were imaged at the indicated time points after being diluted to OD₆₀₀=0.2. Arrowheads, DsRed signal contained within GFP-Pho8 demarcated zones inside vacuoles. d, e Atg8-PE conjugation and deconjugation are dispensable in EVT. GFP-Pho8 expressing cells with the indicated genotype and Atg8 variants were treated as in Fig. 1a. d Fluorescent microscopy images assessing the accumulation of intravacuolar structures. e Immuno-blots assessing the proteolytic processing of GFP-Pho8. See Fig. S2C for quantification