Figure 7

Role of Rfs in the mitochondrial membrane potential, reactive oxygen species generation, and M. lusitanicus virulence. Germinules formed at 3 h of growth in YNB with 0.1% glucose (YNB-0.1%) were visualized (A) in the presence of MitoTracker Green FM, following which the cells were visualized by confocal microscopy (120 ×), bars = 20 mm. The fluorescence signal was quantified using the (B) Mitotracker signal and (C) hydroxyl radical (OH⁻) concentration. The results presented are the average of values obtained from four independent experiments. Significance testing was performed using ANOVA with Fisher's exact test, *P < 0.05; **P < 0.01; ***P < 0.001. (D) Effect of 10 mM N- acetylcysteine (N-ace) or (E) 0.5 mM potassium cyanide (KCN) on the virulence of cell-free supernatants of the culture (SS) from MU636-, R7B-, and rfs-overexpression (MU636 + rfswt) strains aerobically grown (H-SS) in YNB-0.1%. The SSs were used to treat Caenorhabditis elegans. A total of 15–20 nematodes were used per experiment and incubated at 18 °C for 72 h. The results presented are the average of values obtained from four independent experiments. Data were statistically analyzed using the Mantel-Cox test. ***P < 0.01. When results were not considered significant, we did not provide an additional indication (P > 0.05).