Fig. 2

Infant mice release more NETs than adult mice. a Bone marrow-isolated neutrophils from mice were cultured with PMA and the production of ROS determined. b Neutrophils were cultured with LPS (10 ng/ml) for 0, 3, and 6 h and the production of nitric oxide measured. c, d Neutrophils were stimulated in vitro with PMA (100 ng/ml) (c) or LPS (500 ng/ml) (d) for 4 h. Concentrations of cfDNA/NETs (NETs) in the culture supernatant were determined. Mice were injected with LPS (e) or microbial suspension (sepsis) (f). NET concentration in the serum was determined 6 h or 18 h later, respectively. Neutrophils were cultured with PMA (g) or LPS (h) for 4 h and the expression of Padi4 mRNA in the cells were determined by qPCR. i Mice were injected with LPS and neutrophils in the peripheral blood collected 18 h later and the expression of Padi4 mRNA was determined. Ctr = control group, Sep = sepsis group. Data are the mean ± SEM, n = 5–6 per group and representative of two experiments, *p < 0.05, **p < 0.01, ***p < 0.001 (one-way ANOVA, Bonferroni’s)