Figure 2

Effect of TIRAP MyD88, TRIF and TRAM deficiency on HMGB1 complex stimulation. Peritoneal macrophages from wild-type, MyD88^−/−, TIRAP^−/−, TRIF^−/− or TRAM^−/− mice were stimulated with preformed HMGB1-LPS (A) or HMGB1-Pam₃CSK₄ (B) complexes or individual components separately. Stimulations with 1 µg/mL LPS or 10 µg/mL Pam₃CSK₄ were used as positive controls. The results of one representative experiment out of three to seven individual experiments performed for each stimulation are shown: wild-type (n = 7), TRAM^−/− (n = 3), TIRAP^−/− (n = 4), TRIF^−/− (n = 3) and MyD88^−/− (n = 3). All values are expressed as mean ± SD. ND, not detected; (A) , unstimulated; □, LPS (1 ng/mL);, HMGB1 (1 µg/mL); ■, HMGB1 (1 µg/mL) + LPS (1 ng/mL); , LPS (1 µg/mL); (B) , unstimulated; □, Pam₃CSK₄ (10 ng/mL); , HMGB1 (20 µg/mL); ■ HMGB1 (20 µg/mL) + Pam₃CSK₄ (10 ng/mL); , Pam₃CSK₄ (10 µg/mL).