Fig 1: Cdo1 tethers Camkk2 to AMPK by interacting with both of them, thereby promoting AMPK phosphorylation.a Primary hepatocytes were infected with AD-Flag-Cdo1, and immunoprecipitation was performed using anti-Flag beads and followed by western blotting with antibodies as indicated (n = 3 independent biological replicates). WCL, whole cell lysate. b, c Hepa1-6 cells were co-transfected with plasmids encoding Flag-Cdo1WT or Flag-Cdo1Y157F and HA-Camkk2, and followed by confocal analyses. Scale bars, 10μm. d Cell lysates from primary hepatocytes was immunoprecipitated with antibody against Camkk2, followed by western blotting (n = 3 independent biological replicates). e Primary hepatocytes were infected with AD-Cdo1WT or AD-Cdo1Y157F, and cell lysates were immunoprecipitated with antibody against Camkk2 and followed by western blotting (n = 3 independent biological replicates). f Quantification of immunoprecipitation (IP) results of e. g Cdo1flox/flox loci-containing primary hepatocytes were infected with AD-Cre for the ablation of Cdo1. Cell lysates were immunoprecipitated with antibody against Camkk2 and followed by western blotting (n = 3 independent biological replicates, and representative blot was shown). h Quantification of IP results of g. i Cell free kinase assay with purified recombinant Cdo1, Camkk2 and AMPK proteins. The mixtures were detected by western blotting with antibodies as indicated (n = 3 independent biological replicates, and representative blot was shown). j Quantification of western blotting results of i. k Primary hepatocytes were infected with AD-Cdo1 and AD-shCamkk2 for 48 h. Cell lysates were then analyzed by western blotting (n = 3 independent biological replicates, and representative blot was shown). l Quantification of western blotting results of k. m Primary hepatocytes were infected with AD-Cdo1 and AD-shLKB1 for 48 h. Cell lysates were then analyzed by western blotting (n = 3 independent biological replicates, and representative blot was shown). n Quantification of western blotting results of m. Experiments were performed 3 times and similar results were obtained in a–e, g, i, k and m. One-way analysis of variance plus Tukey’s post hoc tests was performed in j; two-way analysis of variance plus Tukey’s post hoc tests were performed in f, h, l and n. All data show the means ± SD. Source data are provided as a Source Data file.