Distinct Redox Signalling following Macrophage Activation Influences Profibrotic Activity

Figure 4

Effect of NOX2 siRNA on M1 and M2 macrophage-derived superoxide. Knockdown of NOX2 mRNA expression in THP-1 macrophages using NOX2 siRNA was confirmed in macrophages treated with both IFN-γ+LPS (a) and IL-4 (d) macrophages for 72 hours via RT-qPCR and expressed relative to the average MΦ (untreated) value. Effect of NOX2 siRNA and missense siRNA on the PDBu-stimulated superoxide signal in M(IFN-γ/LPS) and M(IL-4) macrophages detected via L-012-enhanced chemiluminescence. (b, e) Average recordings demonstrating initial background readings (1-30 min), basal superoxide as detected following the addition of L-012 (100 μM; 31-60 min), and PDBu (10 μM)-stimulated superoxide generation (61-120 min) measured as luminescence intensity in arbitrary units (a.u) in M(IFN-γ/LPS) (b) and M(IL-4) (e) macrophages. (c, f) Peak PDBu-stimulated (basal signal subtracted) superoxide generation in M(IFN-γ+LPS) (c) and M(IL-4) (f) macrophages. All results presented as , . , vs. MΦ (1-way repeated measures ANOVA followed by Dunnett’s post hoc test).