Figure 3.

Exosomes do not affect PMA-induced NETs and IL-17 release by neutrophils. (A) Representative images of NET formation by neutrophils PMA stimulation, in the presence or absence of different concentrations of exosomes. Cells were stained for nuclei (DAPI, blue) and H3 Citrullinate R2/8/17 (green). Scale bar, 25 µm. (B) Quantification of NET release by neutrophils pooled from three independent experiments performed using neutrophils from three unique donors. At least 100 cells were quantified from each experiment. The neutrophils used in these experiments were from different donors than those in Figures 1, 2 and 4. Data shown as mean ± SEM (n = 3). ***P < 0.0001, two-way ANOVA. (C) mRNA expression levels of il17a relative to hprt1 in neutrophils upon PMA stimulation, in the presence or absence of different concentrations of exosomes (n = 5). Five independent experiments were performed using neutrophils from five unique donors. The neutrophils used in these experiments were from different donors than those in Figures 1, 2 and 4. *P = 0.03, unpaired two-tailed Student's t test. (D) Neutrophils were stimulated with PMA in the presence or absence of different concentrations of exosomes, and culture supernatants were assayed for IL-17 by ELISA (n = 3). Three independent experiments were performed using neutrophils from three unique donors. **P = 0.01, unpaired two-tailed Student's t test. (Color version of figure is available online.)