Fig. 9

AMPK activation is independent of classical pathways. A The structure of honokiol and the synthesis of biotin-linked honokiol. B Schematic showing the construction of plasmids expressing 3X-FLAG-PRKAG1, 3X-FLAG-PRKAG2, and 3X-FLAG-PRKAG1-3A, respectively. C Representative gels showing the binding of honokiol to Flag-AMPKγ1 in 293 T cells. n = 3 replicates. D Representative gels showing the binding of honokiol to Flag-labeled AMPKγ1 and Flag-labeled AMPKγ2. n = 3 replicates. E Representative gels showing the binding of honokiol to Flag-labeled AMPKγ1 and Flag-labeled AMPKγ1-3A. n = 3 replicates. F Western blots of p-AMPKα, AMPKα, p-ACC, and ACC of shRNA hepatocytes or shPRKAG1 hepatocytes at the indicated conditions. n = 3 replicates. G Representative images (left) of BODIPY staining and quantification (right) of lipid droplets in shRNA hepatocytes or shPRKAG1 hepatocytes challenged with PO for 12 h under the indicated conditions. n = 3 replicates. One-way ANOVA was used for statistical analysis. Scale bar 10 μm. H Western blots of p-AMPKα, AMPKα, p-ACC, and ACC of shRNA hepatocytes transfected with empty vector and shPRKAG1 hepatocytes transfected with Flag-PRKAG1 or Flag-PRKAG1-3A plasmids and challenged with PA for 18 h under the indicated conditions. n = 3 replicates. I Representative images (left) of BODIPY staining and quantification (right) of lipid droplets in shPRKAG1 L02 hepatocytes transfected with empty vector, Flag-PRKAG1, or Flag-PRKAG1-3A plasmids and challenged with PO for 12 h under the indicated conditions. n = 3 replicates. Scale bar 10 μm. J Schematic showing the mechanism of honokiol-mediated activation of the AMPK complex and beneficial effects of honokiol in ameliorating obesity and NASH progression