Figure 3
Figure 3. ZV50 describes the activation state of adhered platelets in flowing human blood. Fibrillar collagen, monomeric collagen, or combinations of GFOGER, VWF-III, and one of a panel of CRP-like peptides containing increasing numbers of GPO triplets were used to coat slides. These were perfused with DiOC6-labeled whole human blood for 5 minutes at a shear rate of 1000s−1 with or without preincubation with 20 μg/mL of the anti-GpVI 10B12 or 10μM of the αIIbβ3 inhibitory compound, GR144053. (A) Schematic diagram showing the derivation of ZV50 and mean thrombus height. Consecutive volumes of each slice were accumulated sequentially until a value of 50% of the total volume was reached. This value corresponds to ZV50. Mean thrombus height is the thrombus volume per unit area of coverslip imaged (supplemental Figure 1). (B-C) Thrombus formation onto peptide mixtures of GFOGER and VWF-III with one of a panel of CRP-like peptides after treatment with 10B12. (D-E) Thrombus formation onto peptide combinations after treatment with GR144053. (B,D) Surface coverage (%). (C,E) ZV50 (μm). (F) ZV50 values obtained for combinations of CRP, GFOGER, and VWF-III peptides after blood flow at different shear rates. Data are mean ± SEM. Shear rates: indicates 100s−1; , 300s−1; , 1000s−1; and ▭, 3000s−1.

ZV50 describes the activation state of adhered platelets in flowing human blood. Fibrillar collagen, monomeric collagen, or combinations of GFOGER, VWF-III, and one of a panel of CRP-like peptides containing increasing numbers of GPO triplets were used to coat slides. These were perfused with DiOC6-labeled whole human blood for 5 minutes at a shear rate of 1000s−1 with or without preincubation with 20 μg/mL of the anti-GpVI 10B12 or 10μM of the αIIbβ3 inhibitory compound, GR144053. (A) Schematic diagram showing the derivation of ZV50 and mean thrombus height. Consecutive volumes of each slice were accumulated sequentially until a value of 50% of the total volume was reached. This value corresponds to ZV50. Mean thrombus height is the thrombus volume per unit area of coverslip imaged (supplemental Figure 1). (B-C) Thrombus formation onto peptide mixtures of GFOGER and VWF-III with one of a panel of CRP-like peptides after treatment with 10B12. (D-E) Thrombus formation onto peptide combinations after treatment with GR144053. (B,D) Surface coverage (%). (C,E) ZV50 (μm). (F) ZV50 values obtained for combinations of CRP, GFOGER, and VWF-III peptides after blood flow at different shear rates. Data are mean ± SEM. Shear rates: indicates 100s−1; , 300s−1; , 1000s−1; and ▭, 3000s−1.

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