Figure 3.
Pulse–chase analysis of vacuolar hydrolases. Wild-type cells, vps41Δ1 deletion mutants, and vps41Δ1 cells complemented with VPS41 (pWS24) were grown to midlog phase at 30°C, pulse-labeled with Trans 35S-label for 10 min, and chased with excess unlabeled cysteine and methionine, and cells were harvested 0 min and 40 min after the addition of the chase medium. The samples were then separated into intracellular (I) and extracellular (E) fractions, and specific proteins were immunoprecipitated and applied to SDS/PAGE and autoradiography.