Histology
Paraffin-embedded liver samples were cut into approximately 4 μm
sections and stained with haematoxylin and eosin (HE) and Sirius red.
For Sirius red staining, deparaffinized sections were incubated for 15
min with Picrosirius red solution prepared by mixing 5 mL of 1% Sirius
red solution (Muto Pure Chem. Co., Ltd., Tokyo, Japan) with 100 mL of
saturated picric acid solution (Fujifilm Wako Pure Chem. Corp). The
NAFLD activity score (NAS) of each slide was evaluated as previously
described (Kleiner et al., 2005).