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. 2020 Jul 2;10(18):8080–8097. doi: 10.7150/thno.43473

Figure 8.

Figure 8

Flubendazole elicits anti-cancer effects via targeting EVA1A-modulated autophagy and apoptosis in TNBC cells. (A) MDA-MB-231 and MDA-MB-468 cells were transfected with negative-control or EVA1A siRNA, respectively. After treatment with or without flubendazole (0.5 µM) for 24 h, cell viability was measured by MTT assay. (B) MDA-MB-231 and MDA-MB-468 cells were transfected with negative-control or EVA1A siRNA, respectively. After treatment with or without flubendazole (0.5 M) for 2 weeks. Representative images and quantification of colonies were shown. (C) MDA-MB-231 and MDA-MB-468 cells were transfected with negative-control or EVA1A siRNA, followed by treatment with or without flubendazole (0.5 µM) for 24 h. Then, the expression levels of EVA1A, p62, LC3, Bax and Bcl-2 were determined by immunoblotting analysis. β-actin was measured as a loading control. (D-E) MDA-MB-231 and MDA-MB-468 cells were transfected with negative-control or EVA1A siRNA, followed by treatment with or without flubendazole (0.5 µM) for 24 h. Representative images with quantification of LC3 intensity were shown. Scale bar, 10 µm. (F) MDA-MB-231 and MDA-MB-468 cells were co-transfected with EVA1A siRNA and Flag-EVA1A or vehicle control respectively for 24 h, followed by treatment with or without flubendazole (0.5 µM) for 24 h. Immunoblotting analysis of ATG5, EVA1A and LC3 expression. β-actin was measured as a loading control. Quantification of immunoblotting were shown. (G-H) MDA-MB-231 and MDA-MB-468 cells were co-transfected with EVA1A siRNA and Flag-EVA1A or vehicle control respectively for 24 h, followed by treatment with or without flubendazole (0.5 µM) for 24 h. Representative images with quantification of LC3 intensity were shown. Scale bar, 10 µm. (I) MDA-MB-231 and MDA-MB-468 cells were co-transfected with EVA1A siRNA and Flag-EVA1A or vehicle control respectively for 24 h, followed by treatment with or without flubendazole (0.5 µM) for 24 h, cell viability was measured by MTT assay. (J) MDA-MB-231 and MDA-MB-468 cells were co-transfected with EVA1A siRNA and Flag-EVA1A or vehicle control respectively for 24 h. After treatment with or without flubendazole (0.5 µM) for 2 weeks. Representative images and quantification of colonies were shown. Data are expressed as mean ± SEM. All data were representative of at least three independent experiments. *, P < 0.05, **, P < 0.01, ***, P < 0.001. Statistical significance compared with respective control groups.