Figure 6.

Pho86p is required for packaging of Pho84p-HA, but not Vph1p, into COPII vesicles. Vesicle budding experiments and quantitation were performed as in Fig. 5. Permeabilized spheroplasts prepared from a pho86Δ mutant expressing Pho84p-HA (EY0668) were utilized in this experiment. Pho84p-HA was immunoprecipitated from both total and vesicle fractions by using anti-HA antibodies. Vph1p was then immunoprecipitated by using anti-Vph1 antibodies. Both proteins were resolved separately on 8% SDS/PAGE and analyzed with a PhosphorImager.