Fig. 2

NOX2 deficiency alters macrophage response to LPS/IL-4. LPS/IL-4 (both 10 ng/ml; 24 h) stimulation significantly increased the release of TNFα (a), IL-6 (b), and Nitrite (c) in WT and NOX2^−/− BMDMs (^*** P < 0.001; ANOVA). LPS/IL-4-induced increase in pro-inflammatory markers was significantly reduced in and NOX2^−/− BMDMs compared with WT BMDMs (⁺⁺ P < 0.01, ⁺⁺⁺ P < 0.001, WT LPS/IL-4 vs. NOX2^−/− LPS/IL-4; ANOVA). LPS/IL-4 increased Arg1 (d) protein expression (representative western immunoblot shown (e)) and YM1 (f) mRNA expression in WT and NOX2^−/− (^*** P < 0.001; ANOVA). Expression of anti-inflammatory markers was significantly increased in NOX2^−/− BMDMs compared with WT BMDMs (⁺⁺ P < 0.01, ⁺⁺⁺ P < 0.001, WT LPS/IL-4 vs. NOX2^−/− LPS/IL-4; ANOVA). LPS/IL-4 significantly decreased CD206 mRNA expression in WT BMDMs (^* P < 0.05; ANOVA; g), while CD206 mRNA expression was significantly increased in NOX2^−/− BMDMs following LPS/IL-4 stimulation (⁺⁺⁺ P < 0.001, NOX2^−/− con vs. NOX2^−/− LPS/IL-4; ANOVA). All data are expressed as means (±SEM; n = 6)