Figure 1.

Effect of metformin on nuclear factor-κB (NF-κB) signaling and gene expression. A–C, Primary hepatocytes were incubated in serum-free medium overnight and then stimulated for 3 h with or without 0.5 to 5 mmol/L metformin. For the last 15 min, cells were treated with or without 10 ng/mL tumor necrosis factor (TNF)-α. Cells were lysed and prepared for immunoblotting using antibodies as described in the Methods section of this article. In this figure and elsewhere, each blot is representative of experiments carried out at least 3×. D–F, Primary hepatocytes were incubated as in A–C, before stimulation for 3 h with or without 2 mmol/L metformin and TNF-α. In addition, cells were incubated with/without 10 μmol/L BI605906 or 100 nmol/L rapamycin as shown, before lysis and immunoblotting as described in the Methods section of this article. G–J, Primary hepatocytes were treated with or without 10 ng/mL TNF-α, 2 mmol/L metformin, or 10 μmol/L BI605906 for 8 h followed by cell lysis, RNA extraction, and preparation of cDNA for real-time-polymerase chain reaction using primer sets for individual genes shown as described in the Methods section of this article. ACC indicates acetyl-CoA carboxylase; AMPK, AMP-activated protein kinase; p-ACC, phospho–acetyl-CoA carboxylase; and pAMPK, phospho–AMP-activated protein kinase.