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. 2000 Mar;11(3):969–982. doi: 10.1091/mbc.11.3.969

Figure 1.

Figure 1

Epitope-tagged Apg5p complements defects in prAPI import and macroautophagy in an apg5Δ strain. (A) API maturation in steady-state cultures. Cells from strains SEY6210 (WT, lane 1), apg5 (lane 2), apg5Δ (lane 3), and apg5Δ expressing Apg5HAp from a CEN plasmid (HA, lane 4) were grown to log phase, and protein extracts were prepared and immunoblotted with antiserum to API, as described in MATERIALS AND METHODS. mAPI, mature API. (B) Kinetic analysis of prAPI import. Cells from strains SEY6210 (WT), apg5Δ, and apg5Δ expressing Apg5HAp from a CEN plasmid (APG5-HA) were pulse labeled for 10 min at 30°C and subjected to a nonradioactive chase for the indicated times. API was recovered by immunoprecipitation and detected with a STORM PhosphorImager. The percentage of mature API (%mAPI) was calculated by dividing mature API by mature API plus prAPI at each time point. (C) Viability during nitrogen starvation. Cells from SEY6210 (WT), apg5Δ, and apg5Δ expressing Apg5HAp from a CEN plasmid (APG5-HA) were analyzed for sensitivity to starvation, as described in MATERIALS AND METHODS.