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. 2022 Jan 19;13:386. doi: 10.1038/s41467-022-28044-x

Fig. 4. HADHA and BHB inhibited glucagon-induced FOXO1 activation.

Fig. 4

a Luciferase reporter assay of the inhibitory effect of HADHA on Pck1 and G6pc gene promoter co-transfected with HADHA and FOXO1 plasmid in HEK-293T cells. The luciferase activity was normalized with the internal control (Renilla luciferase, n = 6). b Representative confocal images of FOXO1 nuclear translocation in HADHA-overexpressed primary hepatocytes with or without BDH1 siRNA transfection and 100 nM glucagon stimulation for 1 h. Scale bar represents 10 μm. c Confocal images of FOXO1 nuclear translocation in BHB (400 μM, 6 h) treated primary hepatocytes when exposed to glucagon (100 nM, 1 h). Scale bar represents 10 μm. d FOXO1 phosphorylation from the hepatocytes in b (n = 3). e FOXO1 phosphorylation from the hepatocytes in panel c (n = 3). f, g FOXO1 acetylation in HADHA-overexpressed or knockdown primary hepatocytes transfected with BDH1 siRNA and 100 nM glucagon stimulation for 1 h. b, c, f and g were repeated 3 times independently with similar results. BHB β-hydroxybutyrate, GLC glucagon, NC normal control. Values represent mean ± SEM. Statistical differences were determined by one-way ANOVA. Source data are provided as a Source Data file.