Figure 2.

Vacuolar morphology is unaltered in the Vam3ΔN mutant. Wild-type or mutant BJ3505 strains were incubated with 10 μM of the dye FM4–64 (Molecular Probes, Eugene, OR) for 20 min at 30°C in YPD. Cells were centrifuged briefly (1 min at 5000 × g), washed twice with YPD medium, and chased for 15 min at 30°C (Vida and Emr, 1995). Stained vacuoles were analyzed in a standard fluorescence microscope. (A and B) BJ3505 wild-type cells; (C and D) BJ3505 VAM3ΔN cells; (E and F) BJ3505 vam3Δ cells as a negative control.