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Fig. 5 | Journal of Neuroinflammation

Fig. 5

From: NOX2 deficiency alters macrophage phenotype through an IL-10/STAT3 dependent mechanism: implications for traumatic brain injury

Fig. 5

Increased anti-inflammatory activation in NOX2−/− BMDMs is mediated through IL-10. (a) Representative immunoblots for pSTAT3, STAT3, Arg1, pSTAT6, STAT6, and β-actin. (b) LPS/IL-4 (both 10 ng/ml; 24 h) increased the expression of pSTAT3 in WT BMDMs (* P < 0.05, vs. control; ANOVA), and LPS/IL-4-induced pSTAT3 expression was significantly increased in NOX2−/− BMDMs (++ P < 0.01, WT LPS/IL-4 vs. NOX2−/− LPS/IL-4; ANOVA). Co-incubation of BMDMs with neutralizing anti-IL-10 (αIL-10) attenuated the LPS/IL-4-induced increase in pSTAT3 in NOX2−/− BMDMs (^^^ P < 0.001, NOX2-/- LPS/IL-4 vs. NOX2-/- LPS/IL-4 + αIL-10; ANOVA). (c) αIL-10 attenuated the LPS/IL-4-induced increase pSTAT6 expression (*** P < 0.001, control vs. LPS/IL-4; ## P < 0.01, WT LPS/IL-4 vs. WT LPS/IL-4 + αIL-10; ^^^ P < 0.001, NOX2−/− LPS/IL-4 vs. NOX2−/− LPS/IL-4 + αIL-10; ANOVA), and no genotype-related changes were observed. (d, e) LPS/IL-4 induced an increase in Arg1 (d) protein expression, and IL-4Rα (e) mRNA expression in WT and NOX2−/− BMDMs (*** P < 0.001, vs. control; ANOVA); these effects were significantly increased in NOX2−/− BMDMs (+ P < 0.05, +++ P < 0.001, WT LPS/IL-4 vs. NOX2−/− LPS/IL-4; ANOVA). The LPS/IL-4 mediated effects on Arg1 and IL-4Rα expression were significantly reduced in the presence of αIL-10 (^^^p < 0.001, NOX2−/− LPS/IL-4 vs. NOX2−/− LPS/IL-4; ###p < 0.001, WT LPS/IL-4 vs. WT LPS/IL-4 + αIL-10; ANOVA). (f, g) SOCS3 and IL-10 mRNA expression was significantly increased in WT and NOX2−/− BMDMs following LPS/IL-4 stimulation (* P < 0.05, *** P < 0.001, vs. control; ANOVA); the LPS/IL-4 effect was significantly increased in NOX2−/− BMDMs compared with WT BMDMs (* P < 0.05, WT LPS/IL-4 vs. NOX2−/− LPS/IL-4; ANOVA). SOCS3 and IL-10 expression levels were significantly increased in the presence of αIL-10 (^^ P < 0.01, ^^^ P < 0.001, NOX2−/− LPS/IL-4 vs. NOX2−/− LPS/IL-4 + αIL-10; ### P < 0.001, WT LPS/IL-4 vs. WT LPS/IL-4 + αIL-10; ANOVA). All data are expressed as means (± SEM, n = 6)

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