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Blood, Vol. 111, Issue 1, 200-208, January 1, 2008
The factor VIII C1 domain contributes to platelet binding
Blood Hsu et al.
111: 200
Supplemental material for: Hsu et al
Files in this Data Supplement:
- Figure S1 A and B. Competition Assays. Competition of fluorescein-labeled C1C2-2296C (C1C2*) by unlabeled FVIIIa (PDF, 71.9 KB). -
A. Histogram corresponding to one of the six competition experiments summarized in Figure 6C (left panel). The grey peak on the far left shows counts from activated platelets only, measured as a control. The broad peak enclosed by a bold solid line shows platelets after addition of 0.25 µM C1C2*. The peaks enclosed by light solid and dotted lines show activated platelets after addition of 0.25 µM C1C2*, followed by FVIIIa at molar ratios of FVIIIa:C1C2* = 1:1 and 4:1, respectively. The broadness of the peaks varied somewhat among different platelet preparations, but the competition trends were quite similar. The histogram for FVIIIa:C1C2* = 2:1 was omitted to avoid crowding the figure. B. Scattergram representation of the same data shown in A, including FVIIIa:C1C2* = 2:1. The measurement of platelets alone is not shown, but was quite similar to the platelets-only results shown in Fig. 4, top panel).
- Figure S1 C and D. Competition Assays. Reciprocal experiments showing competition of fluorescein-labeled FVIIIa (FVIIIa*) by unlabeled C1C2 (PDF, 128 KB). -
C. Histogram representation of data from one of the five experiments summarized in Figure 6D (central panel). The grey peak on the far left shows counts from activated platelets only, measured as a control. The broad peak enclosed by a bold solid line shows platelets after addition of 10 nM FVIIIa*.The peaks enclosed by light solid and dotted lines show activated platelets after addition of 10 nM FVIIIa*, followed by addition of C1C2 at molar ratios of C1C2:FVIIIa* = 50:1 and 100:1, respectively. The broadness of the peaks varied somewhat among different platelet preparations, but the competition trends were quite similar. The histogram for C1C2:FVIIIa* = 200:1 was omitted to avoid crowding the figure. D. Scattergram representation of the same data in C, including C1C2:FVIIIa* = 200:1. The measurement of platelets alone is not shown, but was 4%, quite similar to the C1C2:FVIIIa* = 200:1 result, demonstrating that C1C2 can compete off essentially all of the FVIIIa* bound to activated platelets under these conditions.
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