Fig. 6
IL-17 effects on the PMA-induced release of NETs. a The mRNA expression of IL-17 in BALF cells before and after one and two weeks of treatment with GC was evaluated by qPCR. Bars represent means ± SEM. * Significant difference from controls at the same time (*p < 0.05, **p < 0.001). b Blood neutrophils were incubated 24 h with or without 100 ng/ml reIL-17, cytocentrifuged and stained with a modified Wright–Giemsa solution, and analyzed using the NETs score. c Representative immunofluorescence images were taken one hour after PMA treatment, staining with Histone H3 (green), MPO (red) and DAPI (blue). Higher-magnification views show NETs formation, demonstrated by the co-localization of Histone H3 (green), MPO (red) and DNA (blue). Scale bar represents 20 μm. d Neutrophils isolated from blood were seeded in the presence of 200 nM of PMA, ± 100 ng/ml reIL-17 and ±10−6 M dexamethasone for 1 hour. Cells were stained with 2.5 μM of DNA-staining dye Sytox Orange. Random images taken by a blinded evaluator and NETs quantification were assessed. Bars represent the fold change, expressed as the mean ± SD relative to the PMA-stimulated condition. * Significant differences from PMA (p < 0.05)