Flubendazole inhibits MDA-MB-231 cells migration through autophagy in vitro. (A-B) MDA-MB-231 cells were treated with flubendazole (0.5 µM) alone or in combination with 3-MA (1 mM) for 24 h, 3-MA was added 1 h before treatment of flubendazole. Then scratch assay and transwell assay were used to measure the migration capabilities of the cells. Representative images and statistics were shown. Scale bar, 100 µm. (C-D) MDA-MB-231 cells were transfected with control or ATG5-siRNA, followed by treatment with or without flubendazole (0.5 µM) for 24 h. Then scratch assay and transwell assay were used to measure the migration capabilities of the cells. Representative images and statistics were shown. Scale bar, 100 µm. (E-F) MDA-MB-231 cells were treated with flubendazole (0.5 µM) alone or in combination with 3-MA (1 mM) for 24 h, 3-MA was added 1 h before treatment of flubendazole. The expression of MMP-2 and E-cadherin were analyzed by immunofluorescence. Scale bar, 20 µm. (G-H) MDA-MB-231 cells were transfected with control or ATG5-siRNA, followed by treatment with or without flubendazole (0.5 µM) for 24 h. The expression of MMP-2 and E-cadherin were analyzed by immunofluorescence. Scale bar, 20 µm. 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.