Extended Data Figure 4.

Effect of chronic metformin (50 mg/kg per day, IP x 30 days) treatment on liver redox, energy charge and expression of gluconeogenic regulators. (a) Chronic metformin treatment increased the liver [GSSG]:[GSH] ratio, (b) but had no effect on the liver [ATP]:[ADP] or (c) [ATP]:[AMP] ratios. (d) The [NADH]:[NAD⁺] and (e) [NADPH]:[NADP⁺] ratios also remained unchanged. (f) Chronic metformin treatment slightly reduced liver [cAMP] levels. (g) Chronic metformin treatment had no effect on the protein levels of major gluconeogenic enzymes in the liver, (h) PEPCK-C protein expression and (i) liver PC protein levels both remaining unaltered. (j) Activated CREB as determined by the ratio of phosphorylated CREB to total CREB levels was decreased. (k) Chronic metformin treatment activated liver AMPK as indicated by the increased ratio of phosphorylated AMPK to total AMPK levels and (l) increased phosphorylation of ACC. Data are mean ± SEM, (n=6 biological replicates). * P<0.05, **P<0.01, ***P<0.001 by unpaired t-test.