Figure 2.
Figure 2. Arrest of flow platelets in whole blood on TNF-α–treated vascular endothelial cells incubated with blocking mAbs directed against TSP-1 or SIRPα. EA endothelial cells were grown onto permanox Lab-Tek 1 chamber slides and stimulated with 25 ng/mL rh TNF-α for 18 hours. Then 40 μg/mL anti-TSP-1 C6.7 and A2.5 or anti-SIRPα SE5A5 mAbs were added or not added (control) to these cells for 10 minutes at 37°C. They were washed with PBS before being placed in the flow chamber. After platelet labeling in PRP with calceine, whole blood was reconstituted and perfused through the chamber at 37°C at a shear rate of 100 seconds–1 for 3 minutes. Platelet adhesion, expressed as the percentage of surface covered with platelets, is the average ± SEM of 10 random fields per coverslip. ***P < .001. This experiment is representative of 3 experiments performed using blood from different donors.

Arrest of flow platelets in whole blood on TNF-α–treated vascular endothelial cells incubated with blocking mAbs directed against TSP-1 or SIRPα. EA endothelial cells were grown onto permanox Lab-Tek 1 chamber slides and stimulated with 25 ng/mL rh TNF-α for 18 hours. Then 40 μg/mL anti-TSP-1 C6.7 and A2.5 or anti-SIRPα SE5A5 mAbs were added or not added (control) to these cells for 10 minutes at 37°C. They were washed with PBS before being placed in the flow chamber. After platelet labeling in PRP with calceine, whole blood was reconstituted and perfused through the chamber at 37°C at a shear rate of 100 seconds–1 for 3 minutes. Platelet adhesion, expressed as the percentage of surface covered with platelets, is the average ± SEM of 10 random fields per coverslip. ***P < .001. This experiment is representative of 3 experiments performed using blood from different donors.

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