Figure 5

Effects on secreted msGFP constructs of deleting or mutating Vps10. (A) S. cerevisiae cells, either expressing wild-type Vps10 (“WT”) or carrying a vps10-104 or vps10Δ allele, were engineered to express msGFP fused to the Ost1 signal sequence (pre-Ost1-msGFP). Exposure times for the fluorescence images were 500 msec. Scale bar, 2 μm. (B) Intracellular and extracellular CPY for the indicated strains was analyzed by SDS-PAGE and immunoblotting as described in Methods. (C) msGFP secretion was analyzed as in Figure 1A for the indicated strains, except that immunoblotting was performed with the quantitative procedure described in Methods. The experiment was done twice, with three replicates each time. Immunoblots from one of the experiments are shown. Error bars represent s.e.m.