Western blotting results showed that insulin receptor (IR), protein kinase B (Akt), and glycogen synthase kinase-3β (GSK3β) phosphorylation levels and PPARα protein expression levels were significantly decreased, and sterol regulator element-binding protein-1c (SREBP-1c) protein expression levels were significantly increased in the liver samples from dairy cows with NAFLD compared with those from control cows (Supplementary Figure 1A and 1B). This evidence concerns the gene AKT1 and metabolic dysfunction-associated steatotic liver disease.