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. 2021 Jan 21;8:617214. doi: 10.3389/fcell.2020.617214

Figure 3.

Figure 3

Effects of DB on vacuolar integrity in C. albicans SC5314 and C. glabrata BG2 planktonic cultures in stationary phase. Stationary cultures of C. albicans SC5314 and C. glabrata BG2 were treated with a 2-fold dilution series of domiphen bromide (DB) from 3.125 μM up to 25 μM for 2.5 h. DMSO background concentration was 2%. Concanamycin A (CMA) was used as a positive control for an elevated vacuolar pH. The ratio of fluorescence intensity of dye BCECF,AM from excitation at 490 nm (I490nm) to fluorescence intensity from excitation at 450 nm (I450nm), both measured at an emission wavelength of 535 nm, was calculated and normalized for the cell culture biomass (OD490nm) to obtain a relative vacuolar pH value of the treated cells. Mean values ± SEM are shown for three biological repeats. Statistical analysis was performed to assess a significant increase in (I490nm/I450nm)/OD490nm upon treatment with DB as compared to DMSO control treatment. A one-way ANOVA and Dunnett's multiple comparison test was applied and significant differences (p < 0.05) are shown in orange.