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.