CPY sorting and Vps10p stability are
defective in ric1Δ and ypt6Δ strains.
(A) CPY sorting. Wild-type (WT, SEY6210),
ric1Δ (GPY1701) and ypt6Δ (GPY1700)
strains were grown and subjected to pulse-chase analysis as described
in the legend to Figure 1B. After a 15-min labeling period, samples
were removed after 0, 10, and 30 min of chase and CPY was
immunoprecipitated from internal (I) and external (E) fractions. CPY
was resolved by SDS-PAGE and subjected to phophorimage analysis. (B)
Vps10p stability. Wild-type (WT, SEY6210),
ric1Δ (GPY1480), and ric1Δ
pep4Δ (GPY1492) strains were grown and subjected to
pulse-chase analysis as described in the legend to Figure 1B. Following
a 10-min labeling period, cells were removed after 0, 60, and 120 min
of chase and Vps10p was immunoprecipitated from cell lysates. Vps10p
was visualized by SDS-PAGE followed by autoradiography. The 170-kDa
Vps10p proteolytic cleavage product is denoted by an asterisk.