Cell-based hemostasis vs. platelet-based hemostasis
http://www.akh-wien.ac.at/biomed-research/htx/platweb1.htm
http://www.akh-wien.ac.at/biomed-research/htx/platweb1.htm
Asimetría de fosfolípidos de la membrana celularActividad procoagulante de las plaquetas
EscramblasaEscramblasa TranslocasaTranslocasa FlopasaFlopasa
ProtrombinaProtrombina
TrombinaTrombina
MacrófagoMacrófago
VesículaVesícula
CalpaínaCalpaína
CitoesqueletCitoesqueletoo
Hemophilia
Coagulation
Figure 1
A
200 bp
600 bp
B
200 bp
600 bp
200 bp
600 bp
C
2000 bp
1500 bp
2000 bp
CD14β-actin
TF GPIbβ-acti
n
CD14TF GPIb
Non-stimulated PBMC
PBMC +TRAP
Non-stimulated platelets
TRAP-stimulated platelets
Non-stimulated PBMC
LPS-stimulated PBMC
β-actin
CD14TF GPIb
β-actinCD14
TF GPIb
β-actin
CD14TF GPIb
β-actin
TF
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
A 60.4 kDa 47.2 kDa
35 kDa
TRAP – – + +Puromycin – + – +
Figure 3-S
7,5 mPearson Coeff. : 0.1855
Pearson Coeff. : 0.3188
Figure 3
7,5 m
TF GPIb Merge
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 kDa47.2 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 kDa47.2 kDa
35.1 kDa
24.9 kDa
Figure 4
B. Platelets
*
1 2 3 4
62.0 kDa
47.5 kDa32.5 kDa
LPS + + + + Actinomycin D – + – – DRB – – + –Puromycin – – – +
Figure 4S
MergeGPIbαTF
3 m
Hoechst Vis
Figure 5
Merge
GPIb
GPIb
TF
TF
VisTF GPIbα
TF GPIbα Merge
Figure 5-S
TFGPIb
Merge
GPIbα TF
Merge
GPIbα TF
Merge
Figure 6
GPIbα TF
Merge
6 μM
0 25 50 75 100 1250
10
20
30 DMSOPuromycin
Time (minutes)
Proc
oagu
lant
act
ivity
(AU
/ 2x
107 p
late
lets
)
0 25 50 75 100 1250
10
20
30 DMSOPuromycin
Time (minutes)
Proc
oagu
lant
act
ivity
(AU
/ 2x
107 p
late
lets
)
Figure 1-S
0 15 60 1200
50
100
150
200
250
Control
aPC
CTI
-FVIII:C-TF
-FVIIa
Time (min)Activation with 5M TRAP
Proc
oagu
lant
Act
ivity
(%)
Figure 2-S
Regular ManuscriptHuman 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)
