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Blood, Vol. 108, Issue 12, 3746-3752, December 1, 2006
P-selectin glycoprotein ligand 1 and  2-integrins cooperate in the adhesion of leukocytes to von Willebrand factor
Blood Pendu et al.
108: 3746
Supplemental materials for: Pendu et al, Vol 108, Issue 12, 3746-3752
Files in this Data Supplement:
- Video S1. Adhesion of resting PMNs on VWF in flow (MOV, 653 KB)
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PMNs were purified from human blood and perfused over VWF-coated coverslips at a density of 5 × 105 cells/mL and a constant wall shear rate of 50 s–1 for 6 minutes. The experiment was digitally recorded in real time with phase-contrast optics using a 10× Hoffman Modulation Contrast objective and a charge-coupled device camera. Original magnification, 100×. The selected video fragment shows events occurring during 24 seconds of the fourth minute. Note that adhesion of resting PMNs to VWF occurs mainly through transient contact. Quantification of transient contact per minute during a 6-minute period is depicted in Figure 2A. Few firmly adherent cells were observed, whether in this field or in various other fields examined after 6 minutes (not shown). Quantification of firmly adherent cells from 8 independent experiments is shown in Figure 2B.
- Video S2. Adhesion of activated PMNs on VWF in flow (MOV, 773 KB)
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PMNs were purified from human blood and activated with phorbol myristate acetate (100 nM) for 15 minutes before 6-minute perfusion over VWF-coated coverslips at a density of 5 × 105 cells/mL and a constant wall shear rate of 50 s–1. The experiment was digitally recorded in real time with phase-contrast optics using a 10× Hoffman Modulation Contrast objective and a charge-coupled device camera. Original magnification, 100×. Events occurring during 18 seconds of the first minute are displayed in the selected video fragment. In this fragment, activated PMNs adhere firmly to the VWF surface, and few cells have transient contacts. When various fields are observed at the end of perfusion, the accumulation of adherent cells can clearly be observed: the VWF surface is covered by PMNs, some of which are already spread. Quantification of firmly adherent cells from 7 independent experiments is shown in Figure 2B.
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