1 Cell-based hemostasis vs. platelet-based hemostasis

2 http://www.akh-wien.ac.at/biomed- research/htx/platweb1.htm

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6 Asimetría de fosfolípidos de la membrana celular Actividad procoagulante de las plaquetas Escramblasa Translocasa Flopasa Protrombina Trombina Macrófago Vesícula Calpaína Citoesquelet o

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8 Hemophilia

9 Coagulation

10 Figure 1 A 200 bp 600 bp B 200 bp 600 bp 200 bp 600 bp C 2000 bp 1500 bp 2000 bp CD14 β-actin TF GPIb  β-actin CD14 TF GPIb  Non-stimulated PBMC PBMC + TRAP Non-stimulated platelets TRAP-stimulated platelets Non-stimulated PBMC LPS-stimulated PBMC β-actin CD14 TF GPIb  β-actin CD14 TF GPIb  β-actin CD14 TF GPIb  β-actin TF

11 1 2 3 4 5 47 kDa A IgG Basal 30 60 90 min min min B 47 kDa 37 kDa 64.2 kDa 83 kDa 1 2 3 4 Basal 30 60 90 min min min 1 2 3 4 5 47 kDa A IgG Basal 30 60 90 min min min B 47 kDa 37 kDa 64.2 kDa 83 kDa 1 2 3 4 Basal 30 60 90 min min min Figure 2 1 2 3 4 5 47 kDa A IgG Basal 30 60 90 min min min B 47 kDa 37 kDa 64.2 kDa 83 kDa 1 2 3 4 Basal 30 60 90 min min min

12 A 60.4 kDa 47.2 kDa 35 kDa TRAP – – + + Puromycin – + – + Figure 3-S

13 7,5  m Pearson Coeff. : 0.1855 Pearson Coeff. : 0.3188 Figure 3 7,5  m TF GPIb  Merge

14 62.0 kDa 47.5 kDa 32.5 kDa 1 2 3 4 TRAP – – + + Puromycin – + – + 1 2 3 4 5 6 7 8 TRAP – + – + – + – + Tunicamycin – – + + – – + + Puromycin – – – – + + – – LPS – + + – + + Puromycin – + – – – + 109. 5 kDa 78.9 kDa 60.4 kDa 47.2 kDa 35.1 kDa 24.9 kDa 109.5 kDa 1 2 3 4 5 6 78.9 kDa 60.4 kDa 47.2 kDa 35.1 kDa 24.9 kDa Figure 4 * A. Platelets B. Platelets C. PBMC suspensions 62.0 kDa 47.5 kDa 32.5 kDa 1 2 3 4 TRAP – – + + Puromycin – + – + 1 2 3 4 5 6 7 8 TRAP – + – + – + – + Tunicamycin – – + + – – + + Puromycin – – – – + + – – LPS – + + – + + Puromycin – + – – – + 109. 5 kDa 78.9 kDa 60.4 kDa 47.2 kDa 35.1 kDa 24.9 kDa 109.5 kDa 1 2 3 4 5 6 78.9 kDa 60.4 kDa 47.2 kDa 35.1 kDa 24.9 kDa Figure 4 B. Platelets *

15 1 2 3 4 62.0 kDa 47.5 kDa 32.5 kDa LPS + + + + Actinomycin D – + – – DRB – – + – Puromycin – – – + Figure 4S

16 Merge GPIbα TF 3  m HoechstVis Figure 5

17 Merge GPIb  TF Vis TFGPIb α TFGPIbα Merge Figure 5-S

18 TF GPIb  Merge GPIbα TF Merge GPIbα TF Merge Figure 6 GPIbα TF Merge 6 μM

19 Figure 1-S

20 Figure 2-S

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22 Regular Manuscript Human platelets synthesize and express functional tissue factor. Olga Panes1, Valeria Matus1, Claudia G. Sáez1, Teresa Quiroga2, Jaime Pereira1, Diego Mezzano1. From the Departments of Hematology-Oncology1, and Clinical Laboratory2, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile. Supported by Grants 8010002 (D.M., J.P., T.Q.) and 1060637 (D.M.) from Fondecyt, Chile. (Blood, 2007, en prensa)