Figure 3: Development of hepatic steatosis in C57BL/6 mice fed chow, cholesterol, or cholesterol supplemented with formononetin or 2-heptyl-formonetin (C7F) for five weeks (Experiment  2). (A) Weight of liver ( ). (B) Triglyceride content in liver ( ). Total lipids were extracted from liver using a modified version of the Bligh and Dyer protocol, and the content of triglyceride were analysed with a commercial kit. (C) Liver sections stained with hematoxylin and eosin. (D) Hepatic gene expression of Tnf (tumour necrosis factor ) measured by RT-PCR. Data is normalised to 18S ribosomal RNA and presented relatively to the expression in chow ( ). (E) Plasma level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ( ). Graphs show mean ± SEM. Different letters (a, b) denote significant difference ( ) between the groups.