Effect of HMGB1 neutralization on APAP (530 mg/kg; 24 h) hepatotoxicity. Mice were treated with an HMGB1-neutralizing antibody or a control IgY antibody 2 h after APAP treatment and (A) percent (%) survival over 24 h (Fed mice + APAP solid line; fasted mice + APAP/IgY, large dashed line; fasted mice + APAP/anti-HMGB1, small dashed line), (B) serum full-length K18 level (pmol/mL), (C) serum ALT activity (U/L), (D) serum total HMGB1 content, (E) serum TNF-α level (pg/mL) and (F) serum IL-6 level (pg/mL) were recorded simultaneously within the same group of mice. Data are representative of mean ± SD of six mice per group carried out in three independent investigations. Statistical significance was assigned between groups as appropriate. *P < 0.05, **P < 0.01 and ***P < 0.005.