Fig. 5: AgApyrase inhibits platelet-mediated NET formation.

a, b Neutrophils were incubated with platelets in the presence or absence of rAgApyrase. NETs were quantified by immunofluorescence microscopy using an anti-MPO (myeloperoxidase) antibody (a). DNA was stained with DAPI. Yellow arrows point released neutrophil DNA stained with MPO. The percentage of NETs (b) was calculated as an average of 5–10 fields (400X) normalized to total number of neutrophils. Results expressed as mean % ± SEM. n = 4. One-way ANOVA with Šídák’s multiple comparisons test, ***P = 0.0006 for control vs 5 µM and 0.001 for 10 µM. c, d AgApyrase reduces neutrophil ROS production. Whole cell (c) or mitochondrial (d) ROS production in neutrophils activated or not with phorbol myristate acetate (PMA) and incubated in the presence or absence of rAgApyrase. One-way ANOVA with Friedman test. **P = 0.003. n = 6. Error bars indicate SEM. e Immunohistochemistry was performed to quantify neutrophil elastase as a marker for NETosis. f Quantification of the percentage area and the mean signal of neutrophil elastase staining per midgut (additional representative IHC images shown in Fig. S10). Data pooled from two independent experiments. Each dot represents an individual midgut. n = 22 and 35 midguts for control and rAgApyrase, respectively. Mann Whitney test, ****P = 0.0001. Source Data are provided as a Source Data file.