Fig. 1: Hepatic Cdo1 is induced through the cAMP/PKA/CREB pathway during exercise in mice.

a Treadmill training in chow diet-fed 8-week-old mice. The representing mouse model was created using BioRender.com. b–e Liver samples were isolated from the male mice after the exercise program shown in a is done. b The liver messenger RNA (mRNA) level of Cdo1 (n = 6 mice per group). c Western blotting of mice liver lysates (n = 3 mice per group). d Quantification of western blotting results of c. e The cAMP level in mice liver (n = 6 mice per group). f CREB enrichment on the indicated gene promoters (n = 6 independent biological replicates). The β-globin gene promoter serves as a negative control. g Schematic representation of Cdo1 proximal promoter constructs used for luciferase assays. The predicted consensus of CREB-binding site is shown in the WT luciferase construct. The red letters indicate mutations of the CREB-binding site in the CREB-Mut construct. h, i Luciferase activities were measured in HEK293T cells. Data were normalized to the vector group (n = 6 independent biological replicates). h Cells were transiently transfected with WT (Wild type) reporter construct as shown in g, along with different amounts of CREB expression vector. i Cells were transiently transfected with WT or CREB-Mut reporter construct, along with control vector or CREB expression vector. j–o Primary mice hepatocytes or HepG2 cells were treated with 10 μM Forskolin for 12 h. j The mRNA level of Cdo1 in primary hepatocytes (n = 6 independent biological replicates). k Western blotting of primary hepatocytes lysates (n = 3 independent biological replicates). l Quantification of western blotting results of k. m The CDO1 mRNA level in HepG2 cells (n = 6 independent biological replicates). n Western blotting of HepG2 lysates (n = 3 independent biological replicates). o Quantification of western blotting results of n. Unpaired two-tailed t tests were performed in b, d, e, j, l, m and o; one-way analysis of variance plus Tukey’s post hoc tests were performed in h and i; two-way analysis of variance plus Tukey’s post hoc tests were performed in f. All data show the means ± SD. Source data are provided as a Source Data file.