*4.3. LPS/D-GalN-Induced ALF Model*

The animal experiments were carried out in compliance with the animal care and use committee guidelines at Gyeongsang National University (GNU-151208-M0068). Male C57BL/6 mice (7 weeks old) were purchased from Koatech Co. (Animal Breeding Center, Pyongtaek, Korea). Animals were kept on a 12 h light/dark cycle in a specific pathogenfree area with food and water freely available in the animal facility for 1 week before the experiment. All experimental animals were randomly separated into five groups as follows: Saline, LPS (1 μg/kg) + D-GalN (400 mg/kg), LPS/D-GalN + YA (10 mg/kg), LPS/D-GalN + YA (50 mg/kg), and LPS/D-GalN + silymarin (25 mg/kg). YA and silymarin were pre-administered for 10 days before LPS/D-GalN by oral gavage. LPS/D-GalN was injected intraperitoneally. Blood and tissues were collected 6 h after LPS/D-GalN injection. Liver tissues were quickly isolated and placed into a deep freezer at −80 ◦C or a 4% paraformaldehyde solution for further experimentation.
