The formalin-fixed liver sections were stained with H & E, Sirius red, and Masson’s trichrome to detect any liver injury and fibrosis. Further, intestine sections were stained with H & E. The liver HE-stained sections were scored using the NAFLD activity score (NAS) system (Brunt et al., 2011 (link)) by our co-author H.-S.L.—an experienced pathologist—and the degree of fibrosis was evaluated by Sirius red stain (Picrosirius Red Stain Kit, Polysciences, Inc., Warrington, PA, United States) and Masson’s trichrome staining (Muto pure chemical co., Ltd.). NIH ImageJ was used to estimate the length and width of gut villi and crypts.
Paraffin-embedded liver sections were stained for F4/80 (ab6640, abcam®) and stained for alpha-smooth actin (a-SMA) (GTX100034, GeneTex). Paraffin-embedded intestine sections were stained for ZO-1 (NBP1-85047, Novus). Standard immunohistochemical (IHC) staining procedures were employed; liver and intestine sections were incubated with a specific primary antibody, as described earlier, followed by incubation with an HRP-linked secondary antibody (Dako, Glostrup, and Denmark) and 3,30-diaminobenzidine (Dako, Glostrup, and Denmark) and were scanned using the NanoZoomer Digital Pathology system.
Paraffin-embedded liver sections were stained for F4/80 (ab6640, abcam®) and stained for alpha-smooth actin (a-SMA) (GTX100034, GeneTex). Paraffin-embedded intestine sections were stained for ZO-1 (NBP1-85047, Novus). Standard immunohistochemical (IHC) staining procedures were employed; liver and intestine sections were incubated with a specific primary antibody, as described earlier, followed by incubation with an HRP-linked secondary antibody (Dako, Glostrup, and Denmark) and 3,30-diaminobenzidine (Dako, Glostrup, and Denmark) and were scanned using the NanoZoomer Digital Pathology system.
Free full text: Click here
