2.3. Oil Red-O Staining

Oil Red-O staining was applied to assess lipid droplet formation in liver tissues according to a method described previously [34]. Briefly, fresh liver tissues were cut into small pieces and frozen immediately with dry ice, the liver tissue was immersed in OCT embedding agent, and was cut into 8-μm sections using a cryostat (Leica, CM3050S, Wentzler, Germany), and then the liver sections were incubated with 0.5 mg/mL of Oil Red-O staining solution for 5 min to show changes in fat accumulation. Hematoxylin staining was performed to visualize the cell nuclei. Finally, the sections were washed with running tap water and mounted with glycerin gelatin.