Fig. 5.

NET extrusion inhibition by in vitro IFX treatment reduces the inflammatory activity of mononuclear cells. To study the effects of infliximab (IFX) on the inflammatory profile of mononuclear cells promoted by NETs, HD neutrophils (n = 3) were treated with TNF-α (8 ng/mL) in the presence or in the absence of IFX (100 mg/ml) for 2 h. Then, supernatants, which were pre-incubated in the presence or absence of 15 U/ml DNAse I for 10 min, were added to PBMCs and further incubated for 24 h. a Nuclear NF-κB protein expression. Representative blot is shown. Lanes from different conditions were run on the same gel but were non-contiguous. Cropping lines are used in the figure. b Relative expression levels of inflammatory genes. a-b Bar graphs show the mean ± SEM. The data were analyzed using a paired-samples t test. *vs. baseline; ^#vs. TNF-α (P < 0.05). AU, arbitrary units; HD, healthy donor; IL, interleukin; NET, neutrophil extracellular traps; NF-κB, nuclear factor-κB; PBMCs, peripheral blood mononuclear cells; STAT, signal transducer and activator of transcription; TNF, tumor necrosis factor