Figure 3

Deficiency of TIRAP, MyD88, TRAM or TRIF does not influence the enhancing effect of HMGB1 complexes on IL-6 production. Peritoneal macrophages from wild-type, TRIF^−/−, TRAM^−/−, TIRAP^−/− or MyD88^−/− mice were stimulated with preformed HMGB1-LPS or HMGB1-Pam₃CSK₄ complexes or individual components separately. Stimulations with 1 µg/mL LPS or 10 µg/mL Pam₃CSK₄ were used as positive controls. Three to seven individual experiments were performed for each stimulation: wild-type (n = 7), TRAM^−/− (n = 3), TIRAP^−/− (n = 4), TRIF^−/− (n = 3) and MyD88^−/− (n = 3). Raw data in each experiment were normalized to 1 µg/mL LPS (A) or 10 µg/mL Pam₃CSK₄ (B) stimulation to generate fold change. All values are expressed as mean ± SD. ND, not detected; ns, not significant. (A) □, LPS (1 ng/mL); ■, HMGB1 (1 µg/mL) + LPS (1 ng/mL); (B) □, Pam₃CSK₄ (10 ng/mL); ■, HMGB1 (20 µg/mL) + Pam₃CSK₄ (10 ng/mL).