Oro reagent

Liver pathological alterations were presented by the established method of our lab (Li et al., 2021 (link)). In brief, liver tissues were fixed, then dehydrated and embedded in paraffin. Paraffin-embedded tissue was cut into 4 μm sections and stained with hematoxylin-eosin (H&E) according to the standard process (Kohypath, Shanghai, China). For Oil Red O (ORO) staining, frozen liver tissues were embedded in Tissue-Tek OCT Compound (Sakura, Tokyo, Japan), cut into ∼8 μm sections, and stained with ORO reagent (Sigma, St. Louis, MO, United States). For immunohistochemical (IHC) analysis, anti-F4/80 (70076 s, cell signaling technology) primary antibody, and biotinylated goat anti-rabbit IgG (BOSTER, SA1022) were applied. Images were captured under a Nikon Eclipse 50i microscope (Nikon, Tokyo, Japan) with a magnification of ×200.

Liver histological changes were examined according to the previously described method (Li Q. et al., 2020 (link)). Briefly, for hematoxylin and eosin (H&E) staining, liver tissues were fixed in 10% formalin solution for 24 h, dehydrated, paraffin-embedded, sectioned to ∼5 μm thickness, and stained with H&E reagent (Kohypath, Shanghai, China). For Oil Red O (ORO) staining, frozen liver tissues were embedded in Tissue-Tek OCT Compound (Sakura, Tokyo, Japan), cut into ∼8 μm frozen sections, and stained with ORO reagent (Sigma, St. Louis, MO, United States). Images were captured under a Nikon Eclipse 50i microscope (Nikon, Tokyo, Japan) with a magnification of 200×.