Oil red o stain kit for cultured

According to the manufacturer’s instructions, collagen deposition was detected via Masson’s trichrome staining (Solarbio, Beijing, China). Images were analyzed using Image J to quantify the fibrotic area-to-total area ratio. An average of five random fields of each section and three random sections of each mouse were used for the analysis. An Oil Red O stain kit (for cultured cells) (Solarbio) was used according to the manufacturer’s instructions. Images were analyzed using Image J to quantify the Oil Red O staining area. Five random detection fields were used, and three independent experiments were conducted.

The TC intracellular content was measured by a total cholesterol assay kit (Applygen, Beijing, China), and the intracellular TC content was calibrated using protein mass. The proteins extracted from the cells were quantified with a Bradford Protein Assay Kit (Bio‐Rad Laboratories, CA, USA). Additionally, intracellular lipid droplets were determined by Oil Red O staining using an Oil Red O stain kit for cultured cells (Solarbio). After washing it with PBS twice, the RAW264.7 cells were fixed with 4% PFA for 20 min and washed for 5 min with 60% isopropanol. Then, they were stained with Oil Red O working solution for 20 min. Finally, any excess dye was washed away with distilled water five times, and the cells were observed under a microscope (Nikon Eclipse C1).