C009 2

The ALT and AST levels in the plasma were measured with assay kits purchased from Nanjing Jiancheng (C009-2, C010-2). The livers were soaked in 4% formaldehyde for 1 day, and subsequently embedded in paraffin for histologic assessment. The prepared slices were further deparaffinized and stained with hematoxylin-eosin staining or Sirius red staining.

Serum alanine aminotransferase (ALT), lactate, and lactic dehydrogenase (LDH) levels were measured with an ALT assay kit (C009-2, Nanjing Jiancheng Bioengineering Institute, Nanjing, China), a lactate assay kit (A019-2, Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and a LDH assay kit (A020-2, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) following the instructions.

The serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in the rat blood were measured with assays purchased from Nanjing Jiancheng (C010-2, C009-2). For the histologic assessment, the livers were fixed in 4% formaldehyde for 24 hours and embedded in paraffin. Liver sections (4μm) were deparaffinized and fixed. The sections were stained with H&E.

For Con-A models, a single dose of Con-A (Sigma-Aldrich, St. Louis, MO) was injected at 15 mg/kg through the tail vein to induce acute hepatitis in mice. Animals were sacrificed to perform analysis from 1–8 h after the injection randomly with random number table. Blood and liver samples were collected.
To knock down the expression of CFB, shRNA-CFB (ordered from Ruizhen, Nanjing) was performed. A solution of shRNA-CFB or vector plasmids was prepared in PBS. Animals were divided into two groups randomly. Mice were injected with shRNA-CFB or vector plasmids by tail vein at 1 mg/kg to induce CFB downregulation. Twenty-four hours after shRNA-CFB injection Con-A was administered intravenously at 15 mg/kg to all animals. After 8 h animals were sacrificed. Blood and liver samples were collected.
Plasma concentrations of alanine transaminase (ALT) were measured using commercially available kits from Nanjing Jiancheng Bioengineering Institute (C009-2).

The AST and ALT levels were measured by assay kits (C010-2, C009-2) purchased from Nanjing Jiancheng (Nanjing, China). For the histologic assessment, the livers were fixed in 4% formaldehyde for 24 h and embedded in paraffin. Liver sections (5 μm) were deparaffinized and fixed. In all experimental groups, the 5-μm-thick sections of the formalin-fixed and paraffin-embedded livers were processed for hematoxylin and eosin staining (H&E) to estimate the degree of hepatic lesions.

The hearts, liver, and kidney were fixed by 4% PFA for overnight, embedded in paraffin or OCT, and sectioned at 7 µm. Slides were stained with Masson's trichrome (ab150686, Abcam) and HE (C0105S, Beyotime) to identify areas of fibrosis and pathology. The levels of aspartate aminotransferase (AST, C010‐2, Nanjing jiancheng) and Alanine aminotranferease (ALT, C009‐2, Nanjing jiancheng) in serum were measured according to the manufacturer's instructions.