FIGURE 4.

Triclabendazole enhances the ROS/JNK/Bax-mitochondrial pathway. The breast cancer cells (MCF-7 or MDA-MB-231) are pre-treated with or without 5 mM NAC (an antioxidant) followed by co-incubation with triclabendazole for 24 h. (A) The ROS levels in MCF-7 and MDA-MB-231 are measured by a DCFH-DA probe. The cells are observed, and representative images are shown. Scale bars: 10 μm, n = 3. (B) Relative ROS levels analyzed by the Leica software in ten randomly chosen fields with each containing ∼40 cells in (A); data are shown as mean ± SD (n = 10). (C) Representative western blots for proteins levels and protein phosphorylation levels including Bax, Bcl-2, JNK, and p-JNK. (D) The mitochondrial membrane potential is determined by the JC-1 assay. The cells are observed by the Leica confocal microscope, and representative images are shown. Scale bars: 10 µm. (E) The cell viability is measured by the CCK-8 assay in MCF-7 and MDA-MB-231 cells with/without NAC treatment, respectively. Statistical analysis is performed by one-way ANOVA with Tukey's multiple comparisons test. ***p < 0.001; ns, not significant; TRI, triclabendazole.