Mkcb3138v

Paraffin-embedded sections were stained with hematoxylin and eosin (200108, Muto Pure Chemicals, Tokyo, Japan) or sirius red (MKCB3138V, Sigma-Aldrich, Tokyo, Japan). For lipid staining, frozen sections were stained with Oil Red O (M3G0644, NACALAI TESQUE, Kyoto, Japan). Macrophages were detected by F4/80 (MCA497R, Bio-Rad, Tokyo, Japan) and VECTASTAIN secondary antibodies (Vector laboratories, Burlingame, USA). To quantify the area of staining by Oil Red O and Sirius Red, images of five random fields from each section were processed with Image J software (National Institute of Mental Health, Bethesda, MD, USA). Each value was expressed as the percentage of the total area of the section. Numbers of F4/80 positive cells were counted and averaged for five random fields of each section.

Paraffin-embedded sections were stained with haematoxylin and eosin (200,108, Muto Pure Chemicals, Tokyo, Japan) or Sirius Red (MKCB3138 V, Sigma–Aldrich, Tokyo, Japan). For lipid staining, frozen sections were stained with Oil Red O (M3G0644, Nacalai Tesque, Kyoto, Japan). Macrophages were detected by F4/80 (MCA497R, Bio–Rad, Tokyo, Japan) and Vectastain secondary antibodies (Vector Laboratories, Burlingame, CA, USA). To quantify the area of staining by Oil Red O and Sirius Red, images of 5 random fields from each section were processed using ImageJ software (National Institute of Mental Health, Bethesda, MD, USA). Each value was expressed as the percentage of the total area of each section that stained positively. The number of F4/80-positive cells was counted and averaged for 5 random fields in each section. The NAS and fibrosis stages were scored by Y.K. in a blinded manner according to the method of Kleiner et al.^{44 (link)}.