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Logarithmic-phase cells of C. glabrata were prepared in SC broth. Vacuolar membranes were first stained with FM4-64. After washing, the pH-sensitive fluorophore BCECF-AM was added to cell suspensions. Note the accumulation of FM4-64 and BCECF-AM within the vacuole lumen of the Δvph2 mutant. Scale bars, 5 μm. The images are representative of three independent replicate experiments.
