Figure 1.

Complement C5b-9 complex induces NET formation and IL-17 release by neutrophils. (A) Flow cytometric analysis of C5b-9 assembly on neutrophils. (B) Representative images of NET formation by neutrophils upon assembly of complements C5b-9. Cells were stained for nuclei (DAPI, blue) and H3 Citrullinate R2/8/17 (green). Scale bar, 25 µm. (C) Quantification of NET release by neutrophils pooled from three independent experiments performed using neutrophils from four unique donors. At least 100 cells were quantified from each experiment. Data shown as mean ± SEM (n = 4). **P = 0.006, two-way ANOVA. (D) mRNA expression levels of il17a relative to hprt1 in neutrophils upon assembly of C5b-9 (n = 3). Three independent experiments were performed using neutrophils from three unique donors. **P = 0.002, unpaired two-tailed Student's t test. (E) C5b-9 were assembled on neutrophils, and culture supernatants were assayed for IL-17 by ELISA (n = 3). Three independent experiments were performed using neutrophils from three unique donors. *P = 0.02, unpaired two-tailed Student's t test. (F) mRNA expression levels of s100a8 and s100a9 relative to hprt1 in neutrophils upon assembly of C5b-9 (n = 4). Four independent experiments were performed using neutrophils from four unique donors. (Color version of figure is available online.)