Fig. 2.

Effects of hinokitiol on respiratory activity. (A, B) C. albicans SC5314 cells were cultured in RPMI 1640 medium with hinokitiol for 10 h. The cells were then stained with CTC, and the fluorescence intensity was measured by flow cytometry. (C) C. albicans SC5314 cells stained with CTC were observed by fluorescence microscopy (scale bars: 100 μm). (D) C. albicans SC5314 cells were cultured in RPMI 1640 medium and treated by 4 μg/mL of hinokitiol for 30 min, 1 h or 2 h at 30 °C. The cells were then stained with CTC, and the fluorescence intensity was detected by flow cytometry. (E, F) Extracted mitochondria were challenged with 2, 4 or 8 μg/mL hinokitiol for 3 h. 100 μM of Fe²⁺ or Fe³⁺ was used in the rescue experiments. Enzyme activity assays of mitochondrial respiratory chains complexes I and II were then carried out. The results of enzyme activities of mitochondrial respiratory chains complexes III and IV were shown in Fig. S8. (G) C. albicans SC5314 cells were treated with 4 μg/mL hinokitiol for 10 h at 30 °C. The transcription levels of genes related to the mitochondrial respiratory chains complex I and II and alternative oxidases were determined by qPCR and are shown as the fold change relative to the control group. (H) C. albicans SC5314 cells were challenged with 4 μg/mL of hinokitiol for 30 min, 1 h or 2 h at 30 °C. The transcription levels of NDH51 and SDH2 were determined by qPCR and were shown as the fold change relative to the control group. The bars represent means ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.