Samples were fixed in 4% paraformaldehyde solution and embedded in paraffin. Then, 5 μm sections were made and mounted on slides for staining with hematoxylin and eosin (H&E, G1120, Solarbio, Beijing, China), Masson (G1340, Solarbio, Beijing, China), Sirius Red (G3632, Solarbio, Beijing, China) and AB-PAS (G1285, Solarbio, Beijing, China), according to the manufactures’ instructions. For Oil Red O staining, frozen sections of the livers were first obtained, and the staining was performed according to the manufacturer’s instructions (G1260, Solarbio, Beijing, China), while the quantification was performed according to a previous protocol [91 (link)].
With H&E staining, hepatocellular steatosis was graded from 0 to 3 based on the percentage of hepatocytes involved (0 = <5%; 1 = 5–33%; 2 = 33–66%; 3 = >66%), according to a previous study [92 (link)]. The NAS score was calculated by steatosis (0–3), lobular inflammation (0−3) and ballooning (0−2), also according to a previous study [32 (link)].
With H&E staining, hepatocellular steatosis was graded from 0 to 3 based on the percentage of hepatocytes involved (0 = <5%; 1 = 5–33%; 2 = 33–66%; 3 = >66%), according to a previous study [92 (link)]. The NAS score was calculated by steatosis (0–3), lobular inflammation (0−3) and ballooning (0−2), also according to a previous study [32 (link)].
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