Fig. 4: AMPK activation promotes mitochondrial fission via INF2.

A, B Parental and INF2-KO HEC-1B cells were treated with DMSO or A-769662 (100 μM) for 6 h, then the cells were stained with DAPI and Mitotracker Orange. Representative confocal images are shown. Scale bar: 10 μm. The mitochondrial lengths were analyzed statistically and shown in (B). Data are shown as means ± SD (n = 30). C, D Parental and INF2-KO HEC-1B cells were treated with DMSO or A-769662 (100 μM) for 6 h, then the cells were stained with DAPI, DRP1, and Mitotracker Orange. Representative confocal images are shown (C). Scale bar: 10 μm. Quantification of DRP1 puncta per mitochondrial length in (D). Data are medians ± interquartile range (n = 30). E Parental and INF2-KO HEC-1B cells were treated with DMSO or A-769662 (100 μM) for 6 h. The cytosol and purified mitochondrial Fractions were isolated and detected by WB analysis with the indicated antibodies. F HEC-1B cells were treated with DMSO or A-769662 (100 μM) for 6 h. The ER Fractions were isolated and detected by WB analysis with the indicated antibodies. P values are calculated using the One-way ANOVA test in (B), and the Kruskal–Wallis test in (D). *p < 0.05, **p < 0.01, ****p < 0.0001, ns: no significant.