Figure 2.

Pex3p and Inp1p segregate to distinct membranous compartments. (A, B) Wild-type BY4742 cells (WT), as well as pex19Δ, pex3-V81E, pex3Δ, and pex19Δ/pex3-V81E mutant cells, were imaged by confocal fluorescence microscopy. Cells expressed Inp1p-GFP and one of Pex3p-mCherry, mCherry-PTS1, Sec13p-mCherry, Pex30p-mCherry, and Rtn1p-mCherry. Maximum intensity projections (MIP) and optical sections (slice) are shown. Bar, 1 μm. (C) PNS from WT, pex3Δ, and pex3-V81E cells was separated by differential centrifugation into 20 and 200 kg supernatant and pellet fractions. Equal amounts of protein were resolved by SDS–PAGE, and immunoblots were probed with antibodies against HA, Pex3p, and G6PDH. (D) Vesicles in PNS prepared from the same strains as in (C) were floated in a step gradient of sucrose solutions of decreasing density. Equal portions of fractions were analysed by immunoblotting for the indicated proteins. Wedge depicts fraction density.

Source data for this figure is available on the online supplementary information page.

Source data for Figure 2 ^{(290.1KB, pdf)}