To determine whether Sirt1 and Sirt6 are causally related to the development of hepatic steatosis and its amelioration by RGZ, and whether their actions are synergistic, we directly inhibited the Sirt1/6 pathway using RNAi-mediated gene silencing targeting Sirt1 and/or Sirt6 in AML12 mouse hepatocytes. This evidence concerns the gene SIRT6 and fatty liver disease.