Fig. 5: Competition between SESN2 and IGF2BP3 regulates HK2 levels, glycolytic flux, and cell proliferation. | Cell Death Discovery

Fig. 5: Competition between SESN2 and IGF2BP3 regulates HK2 levels, glycolytic flux, and cell proliferation.

From: Sestrin2-mediated disassembly of stress granules dampens aerobic glycolysis to overcome glucose starvation

Fig. 5: Competition between SESN2 and IGF2BP3 regulates HK2 levels, glycolytic flux, and cell proliferation.

A Western blot analysis of SESN2, IGF2BP3, and HK2 expression n HepG2 cells after transduction with control (pLKO.1) or independent shRNAs targeting SESN2 comparing cells cultured with (+) or without (−) 4 g/L glucose for 12 h. Actin served as a loading control throughout. B Western blot analysis of SESN2 and HK2 expression in HepG2 cells 24 h after transfection with Flag control or Flag-SESN2 vectors before culture with (+) or without (−) 4 g/L glucose for 12 h. C HepG2 cells were transfected with the indicated combinations of Flag control, Flag-SESN2 and Flag-IGF2BP3-GFP vectors before determining the expression of HK2 protein and mRNA levels by Western blot (left) and qPCR (right), respectively. Transfection efficiency of SESN2 and IGF2BP3 was confirmed by blotting against anti-Flag. D The expression of HK2 protein (left) and mRNA (right) in HepG2 cells transduced with the indicated combinations of control shRNA (pLKO.1) and shRNAs targeting SESN2 and IGF2BP3. E, F Glycolysis stress test profiles measuring ECAR (E) and glucose uptake measured with 2-NBDG (F) in HepG2 cells transfected with Flag control, Flag-SESN2, or Flag-IGF2BP3-GFP vectors for 24 h culture in medium with (+) or without (−) 4 g/L glucose (Glc) for 12 h. G, H Glycolysis stress test profiles measuring ECAR (G) and glucose uptake measured with 2-NBDG (H) in HepG2 cells transduced with the indicated combinations of control shRNA (pLKO.1) and shRNAs targeting SESN2 and IGF2BP3. I Cell growth rates (left) and colony formation (right) of HepG2 cells transfected with the indicated combinations of Flag control, Flag-SESN2 and Flag-IGF2BP3-GFP vectors measured as total cell numbers or colony numbers over 6 days and 10 days, respectively. J Cell growth rates (left) and colony formation (right) of HepG2 cells transduced with the indicated combinations of control shRNA (pLKO.1) and shRNAs targeting SESN2 and IGF2BP3 measured as total cell numbers or colony numbers over 6 days and 10 days, respectively. A–J Data represent three independent experiments. C (right), D (right), I (left), J (left) Data are mean ± SD, n = 3, *p < 0.05; **p < 0.01; ***p < 0.001; ns not significant, two-way ANOVA analysis.

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