FIG 4.

Deletion of the VPS27 gene results in the production of multivesicular bodies (MVB). (A) Time course of FM4-64 uptake in living cells; indicated strains were visualized with a fluorescein isothiocyanate (FITC) filter and merged with differential interference contrast (DIC) Nomarski images at the indicated time points for cell contour and visualization of vacuolar compartments. Bar, 5 μm. (B) Indicated cells were incubated in ASN without glucose with indicated dyes and observed by fluorescence or DIC microscopy. Bar, 5 μm. (C) Indicated strains were visualized with an FITC filter and merged with DIC Nomarski images for cell contour and visualization of the Vps27-YFP. (D) Electron microscopy of wild-type H99, vps27Δ, or vps34Δ strains of C. neoformans. Arrows point to multivesicular bodies (MVBs). Bar, 600 nm.