Fig. 3.

Effects of hinokitiol on mitochondrial function. C. albicans SC5314 cells were cultured in RPMI 1640 medium with hinokitiol for 10 h. The cells were then stained with Rh123 (A, B) for measurement of mtΔψ. The cells were lysed for measurement of intracellular ATP content (C). The cells were further stained with DCFH-DA (D, E) or stained with MitoSOX Red (F, G) to determine the intracellular or mitochondrial ROS contents. The fluorescence intensity of the stained cells was detected by flow cytometry and the Gmean value was calculated. The bars represent means ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. (H) C. albicans SC5314 cells were adjusted to 5 × 10⁴ cells/mL in SC medium containing 5 mM Tu, 5 mM NAC or 0.5 mM DTT. 4 μg/mL of hinokitiol were added to the cultures and incubated at 30 °C for 16 h. Growth was measured by a microplate reader at 600 nm. Cells that were not treated with hinokitiol and reducing agents served as the control. Each point represents the mean of three replicated measurements. The OD₆₀₀ values at the final detection point are used for significant difference analysis. *P < 0.05, **P < 0.01, ***P < 0.001.