Fig. 7.

Hepatic Ago2-deficiency enhances energy expenditure associated with protein synthesis and AMPK activation. a, b Western blot analyses of AMPK expression and activation (a) and mRNA expression of the Tfam-mitochondrial genes (b) in the liver of L-Ago2 WT (n = 5) and L-Ago2 KO (n = 5) fed HFD at 25 weeks of age. c ATP, ADP, and ATP/ADP ratio levels in L-Ago2 WT (n = 5) and L-Ago2 KO (n = 5) mice fed HFD at 25 weeks of age. ATP/ADP ratio levels were independently measured with a distinct procedure from the ATP and ADP assays. d Western blot analysis of total and specific protein levels normalized by 12S-genomic DNA in the liver of L-Ago2 WT (n = 5) and KO (n = 5) mice fed HFD at 30 weeks of age. e Serum albumin levels in L-Ago2 WT (n = 8) and L-Ago2 KO (n = 8) mice fed HFD at 25 weeks of age. f Energy consumption rate measured in primary hepatocytes isolated from L-Ago2 WT (n = 4, n = 4, n = 3 for 0, 2 h, 5 h, respectively) and L-Ago2 KO (n = 4, n = 4, n = 4 for 0, 2 h, 5 h, respectively) mice in the presence of 1 mM metformin. g Effect of Ago2-deficiency on expression of AMPK activation in primary hepatocytes isolated from L-Ago2 WT (n = 8, n = 8, n = 8 for 0, 2 h, 5 h, respectively) and L-Ago2 KO (n = 8, n = 8, n = 8 for 0, 2, 5 h, respectively) mice in the presence of 1 mM metformin. The graphs show the quantification of the results. h Effect of Ago2 on nascent protein synthesis. Primary hepatocytes isolated from L-Ago2 WT and L-Ago2 KO mice were treated with or without 200 μM Phenformin (Phen) or 10 μM Rotenone (Rote) for 5 h (n = 15 for control, n = 9 for Phen, n = 6 for Rote). i A proposed role of hepatic Ago2 in suppression of protein translation, leading to AMPK activation. The graphs show the quantification of the results. Data are shown as the mean ± SEM. *p < 0.05, **p < 0.01