Fig. 7. EVA1A overexpression induces DRP1-mediated mitophagy in MDA-MB-231 and MCF-7 cells.
MDA-MB-231 and MCF-7 cells were co-transfected with DRP1 shRNA and Flag-EVA1A or vehicle control respectively for 48 h. A Immunoblotting of DRP1, EVA1A, p62, Parkin, p-Parkinser65, PINK1, and LC3 expression. β-actin was measured as the loading control. B, C The autophagosomes are labeled by LC3 (green fluorescence) protein, and the mitochondria are labeled by TOM20 (red fluorescence) protein. The number of co-localized LC3 and TOM20 was quantified. Scale bar, 10 µm. D, E Colocalization of Parkin (red fluorescence) protein and TOM20 (green fluorescence) protein in MDA-MB-231 and MCF-7 cells following flubendazole (0.5 μM, 24 h) treatment. The number of co-localized Parkin and TOM20 was quantified. Scale bar, 10 µm. Data represent mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Statistical significance compared with respective control groups (all P-values were obtained by one-way ANOVA).