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. 2010 Jun 3;6(6):e1000939. doi: 10.1371/journal.ppat.1000939

Figure 5. Fluconazole treatment disrupts V-ATPase function.

Figure 5

(A) Vacuolar pH of WT (BY4742) cultures treated with fluconazole at specified concentrations for 6 hours in YPD. (B) Proton pumping rate and ATPase activity of WT (BY4742) cultures treated with or without fluconazole for 6 hours in YPD. Three independent batches of vacuolar vesicles were isolated and used to assay V-ATPase function. Cultures of upc2-1 mutant, WYP361, were treated with fluconazole (100 µg/ml), ergosterol (50 µM) or their combination at OD 0.1. Growth (C, representative of three independent experiments) was assessed at 8 hour post-treatment, and vacuolar pH (D) was assessed at 6 hour post-treatment. (E) WT C. albicans cells (SC5314) were grown in YPD with or without fluconazole for 5 hours. FM4-64 was added to the cultures to stain vacuoles for 30 min. Cells were chased with fresh YPD with or without fluconazole for 20 min followed by quinacrine addition for another 5 min. Fluorescence microscopic images of the cells were taken immediately after washing. Means and standard deviations are plotted. p values of two-tailed t-test are shown.