Figure 3.

PMN from Adamts13 ^−/− mice display normal activation patterns. PMN (10⁶/ml) derived from wildtype (129/Sv/Pas) or Adamts13 ^−/− mice were stimulated as indicated for 4 h. (A) Phagocytosis was determined in absence or presence of PMA (50 ng/ml) or formyl-peptide agonist WKYMVm (8 µg/ml) using PE-labeled polystyrene beads (5.7 × 10⁷ particles/ml). After 4 h PMN were analyzed by flow cytometry. (B) PMN were stimulated as in (A) and after 4 h expression of CD11b and CD62L was measured by flow cytometry gating on Ly-6G⁺ cells. (C) The oxidative burst activity was quantified with the fluorogenic dye DCFH-DA that converts into green fluorescent DCF in the presence of reactive oxygen species. Untreated (medium) or with formyl-peptide agonist WKYMVm (8 µg/ml) stimulated PMN were analyzed in a fluorescence reader over time. (D) Apoptosis of PMN was determined with Nicoletti assay after 24 h in absence or presence of cycloheximide (CHX, 10 µg/ml) and GM-CSF (50 ng/ml). The summarized results of tow independent experiments (129/Sv/Pas and Adamts13 ^−/−, n = 5) plus SD are shown. A one-way ANOVA with Bonferroni’s posttest detects no significant difference (p < 0.05) between PMN from wildtype and Adamts13 ^−/− mice.