Skip to main content
. 2022 Nov 10;13:994496. doi: 10.3389/fimmu.2022.994496

Figure 5.

Figure 5

CD13 crosslinking induces neutrophil extracellular traps (NETs). Neutrophils were left untreated (control) or were incubated with mAb 452 or its Fab fragments for 30 minutes, washed, and stimulated with PMA (20 nM) or IL-8 (10 ng/ml) for 240 minutes, and fluorescence of extracellular DNA was evaluated with SYTOX-Green. (A) Time-course of SYTOX-Green fluorescence in cells stimulated with PMA, mAb 452 (anti-CD13) or non-stimulated. Fluorescence values are the mean ± SEM of 6 independent experiments. Statistical significance was calculated using one-way ANOVA with Tukey’s post hoc test (*p< 0.05, **p< 0.01) (*a: in the PMA curve, values are significantly different from control from 120 minutes onwards). (B) Immunofluorescence images of neutrophils stimulated with PMA, mAb 452 or non-stimulated for 240 min, fixed and incubated with Hoechst 33342® to visualize extracellular DNA. (C, D) NETs release from human neutrophils stimulated for 240 min with mAb 452 (C), or Fab 452 plus F(ab’)2 fragments of anti-mouse Ig (D), PMA, or IL-8, or the combinations shown. Statistical significance was computed through a one-way ANOVA with a Tukey post hoc test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.001.