α9fl/fl LysMCre+ mice display reduced neutrophil mediated platelet aggregation and decreased NETosis. (A) Neutrophils from each genotype and wild-type platelets were stirred in a Chrono-log Whole Blood/Optical Lumi-Aggregometer (model 700-2) before the addition of fMLP. Representative aggregation curve for the fMLP stimulated neutrophils-induced platelet aggregation (left). Quantification (n = 4-5/group) (right). (B) Cathepsin G release as quantified by ELISA from the fMLP-stimulated neutrophils (n = 4 mice/group). (C) NETs assay was performed by stimulating neutrophils with thrombin-activated platelets. Representative microphotographs of NETs stained with PlaNET green (stains extracellular DNA, green) and counterstained with Hoechst (stains nuclei, blue) (left). Boxed region (lower magnification). Red insert in the boxed region is magnified and shown in the microphotographs. Quantification of the percentage of cells releasing NETs (n = 4 mice/group) (right). Value for each mouse represents a mean from 2 fields.