Figure 7
From: Mitochondrial dysfunction induced by bedaquiline as an anti-Toxoplasma alternative
Mitochondrial dysfunction and parasite autophagy due to BDQ treatment. A Reduction of T. gondii ATP by BDQ treatment. Extracellular T. gondii tachyzoites (2 × 106/per sample) were incubated with BDQ (0, 5, 10, or 20 μM) for 4 h at 37 °C. The tachyzoites were then lysed to determine ATP levels using a multi-label reader. B JC-10 probed fluorescence images of BDQ-treated T. gondii tachyzoites. Extracellular parasites (2 × 106/per sample) were suspended with BDQ (0, 5, 10, or 20 μM) for 4 h at 37 °C and then detected using the JC-10 probe. DMSO and CCCP were used as negative and positive controls, respectively. J-Monomers (red) and J-aggregates (green) represent parasites with low and high mitochondrial membrane potentials, respectively. C Mitochondrial membrane potential in BDQ-treated T. gondii tachyzoites. The JC-10 probe fluorescence intensity ratio (RLUJ-aggregates / RLUJ-Monomers) for each group was calculated. D, E Elevation of T. gondii ROS levels by BDQ. Tachyzoites (1 × 105) were treated with BDQ for 16 h (D) or 24 h (E), and then ROS levels were detected using the DCFH-DA probe. F Parasite autophagy due to BDQ treatment. Intracellular T. gondii tachyzoites were treated with BDQ (0, 5, 10, or 20 μM) for 16 h at 37 °C, then the autophagic vacuoles of the parasites were stained with Monodansylcadaverine (MDC) for 30 min in the dark, and the mean value of fluorescence intensity was detected for each group. Data are expressed as means ± SD of three independent experiments. Ns: not significant, ***, P < 0.001, **** P < 0.0001.