Extended Data Fig. 1: HIR occurrence relies on increased ROS levels.
a. The average daily food intake of C57BL/6 J mice was determined over a 3-day period (n = 5/group). b. The body weight of C57BL/6 J mice was measured after 3 days on their respective diets (n = 5/group). c. Serum insulin levels of C57BL/6 J mice were measured by ELISA following a 6-hour fast (n = 5/group). d. Hepatic H2O2 levels of C57BL/6 J mice were quantified using a fluorescence microplate assay (n = 5/ group). e. HGP was assessed by hyperinsulinemic–euglycemic clamps (n = 5/ group). f. M + 6-glucose enrichment in hepatic glycogen was analysed by LC–MS/MS (n = 5/ group). g. Serum M + 3-glucose enrichment in C57BL/6 J mice, measured by LC–MS/MS (n = 5/ group). h. Hepatocyte ROS levels were measured by flow cytometry (n = 3/ group). i. Hepatocyte glycogen M + 6-glucose enrichment was analysed by LC–MS/MS (n = 3/ group). j. Hepatocyte glucose output was measured using a colorimetric assay (n = 3/ group). k-o. C57BL/6 J mice were fed a ND or a HFD for 3 days (n = 5/ group). Blood and liver tissue were obtained. Basal glucose level was measured using a strip-based glucometer following an overnight fast (k). FFA in liver (l) and in blood (m) was analysed by the colorimetric assay. TG in liver (n) and in blood (o) was analysed by the colorimetric assay. p-r. C57BL/6 J mice fed with a ND or HFD were treated daily with 50 mg/kg ABT (i.g.) or 20 mg/kg Etomoxir (i.p.) for 3 consecutive days. The H2O2 levels in the livers were detected using fluorescence microplate (p). GIR (q) and HGP (r) under clamp conditions were detected by LC–MS/MS (n = 5/ group). s. C57BL/6 J mice were fed a ND or a HFD for 3 days (n = 5/ group). And liver tissue was obtained. And the hepatic levels of IL-1β, IL-6, and TNF were measured by ELISA. Data are presented as mean ± S.E.M. Individual data points represent biological replicates. Individual data points represent biological replicates. p values were calculated using one-way ANOVA (d-j and p-r) or unpaired two-tailed Student’s t-test (a-c, k-o, and s).
