Fig. 5: AMPK-mediated phosphorylation of INF2 at Ser1077 promotes EC cell proliferation. | Cell Death & Disease

Fig. 5: AMPK-mediated phosphorylation of INF2 at Ser1077 promotes EC cell proliferation.

From: Phosphorylation of INF2 by AMPK promotes mitochondrial fission and oncogenic function in endometrial cancer

Fig. 5

A WB analysis of the indicated proteins in WCLs from parental or INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant. B, C INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant were stained with DAPI and Mitotracker Orange. Representative confocal images are shown in (B). Scale bar: 10 μm. The mitochondrial lengths were analyzed statistically and shown in (C). Data are medians ± interquartile range (n = 30). D, E INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant were stained with DAPI, DRP1, and Mitotracker Orange. Representative confocal images are shown (D). Scale bar: 10 μm. Quantification of DRP1 puncta per mitochondrial length in (E). Data are shown as means ± SD (n = 30). F The cytosol and purified mitochondrial fractions of INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant were isolated and detected by WB analysis with the indicated antibodies. (G) CCK-8 assays were performed in parental and INF2-KO HEC-1B cells. Data are shown as means ± SD (n = 3). (H, I) Colony formation assays were performed in INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant. The area of clones in (H) was analyzed statistically and shown in (I). Data are shown as means ± SD (n = 3). (J, K) EdU assays were performed in INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant. The proportion of EdU+ positive cells was analyzed statistically and shown in (K). Data are shown as means ± SD (n = 30). L–N 3D sphere formation assays were performed using parental and INF2-KO HEC-1B cells stably expressing EV, INF2-WT, SA, or SE mutant, scale bar: 5×, 200 μm; 20×, 50 μm. The numbers and sizes of cell spheres were analyzed statistically and shown in (M, N). Data are shown as means ± SD (n = 3). P values are calculated using the Kruskal-Wallis test in (C), the one-way ANOVA test in (E, I, K, M, N), and the Two-way ANOVA test in (G). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns no significant.

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