Figure 4.
Figure 4. Minor alterations of G6bhu/hu platelet functional responses. (A) Averaged aggregation and ATP release traces for washed platelets (2 × 108/mL) activated with indicated concentrations of CRP (i) and anti-CLEC-2 antibody (Ab) (ii). Area under the curve (AUC) quantification of (iii) platelet aggregation and (iv) ATP release (n = 5-6 per condition, **P < .01 and *P < .05). (B) Heparin-PPACK-fragmin anticoagulated whole blood was flowed over glass coverslips coated with collagen immediately following collection. (i) Brightfield images were immediately collected, followed by staining with PE-conjugated JON/A, FITC-conjugated P-selectin, and Alexa 647–conjugated Annexin V and fluorescence imaging. Analysis quantifying (ii-vi) thrombus surface area coverage and morphological scores of adhered platelets, (n = 5-6; ***P < .001; **P < .01) and (vii-ix) surface area coverage of fluorescently labeled antibodies (n = 5-6; ***P < .001; *P < .05). (C) Total blood loss to body weight ratio of mice following excision of tail tip (n = 12-16; ***P < .001; **P < .01). All data collection and analysis in panels B and C were completed blinded, mean ± SEM. Scale bar, 10 μm.

Minor alterations of G6bhu/huplatelet functional responses. (A) Averaged aggregation and ATP release traces for washed platelets (2 × 108/mL) activated with indicated concentrations of CRP (i) and anti-CLEC-2 antibody (Ab) (ii). Area under the curve (AUC) quantification of (iii) platelet aggregation and (iv) ATP release (n = 5-6 per condition, **P < .01 and *P < .05). (B) Heparin-PPACK-fragmin anticoagulated whole blood was flowed over glass coverslips coated with collagen immediately following collection. (i) Brightfield images were immediately collected, followed by staining with PE-conjugated JON/A, FITC-conjugated P-selectin, and Alexa 647–conjugated Annexin V and fluorescence imaging. Analysis quantifying (ii-vi) thrombus surface area coverage and morphological scores of adhered platelets, (n = 5-6; ***P < .001; **P < .01) and (vii-ix) surface area coverage of fluorescently labeled antibodies (n = 5-6; ***P < .001; *P < .05). (C) Total blood loss to body weight ratio of mice following excision of tail tip (n = 12-16; ***P < .001; **P < .01). All data collection and analysis in panels B and C were completed blinded, mean ± SEM. Scale bar, 10 μm.

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