American Society of Hematology

Fig. 5.

$Fig. 5. Inhibition of FITC-LMWH binding to dead cells by heparan sulfate, pentosan polysulfate, and heparin of various molecular weight forms. / Necrotic Jurkat cells (induced by Camptothecin and discriminated by FACS analysis, as described in Figure 1) were preincubated with increasing concentrations of these heparin-like ligands—heparan sulfate, pentosan polysulfate, unfractionated heparin (average Mr, 15 kd), and pharmaceutical preparations of depolymerized heparin of various average molecular weights (as listed in “Materials and methods”). After 60 minutes, FITC-LMWH was added at 1 μM, and incubation continued for another 10 minutes. Cells were then labeled with PE–annexin V and 7-AAD, and the RFI of the necrotic population was determined by FACS analysis as in Figure 2. The RFI at each concentration of competitive ligand was expressed as a percentage of the RFI of cells in its absence. (▪-▪), heparan sulfate; (▿-▿), unfractionated heparin; (▵-▵), 7 kd LMWH; (□-□), 6 kd LMWH; (●-●), 4.5 kd LMWH; (○-○), 3.5 kd LMWH; and (▴-▴), pentosan polysulfate.$

Inhibition of FITC-LMWH binding to dead cells by heparan sulfate, pentosan polysulfate, and heparin of various molecular weight forms.

Necrotic Jurkat cells (induced by Camptothecin and discriminated by FACS analysis, as described in Figure 1) were preincubated with increasing concentrations of these heparin-like ligands—heparan sulfate, pentosan polysulfate, unfractionated heparin (average Mr, 15 kd), and pharmaceutical preparations of depolymerized heparin of various average molecular weights (as listed in “Materials and methods”). After 60 minutes, FITC-LMWH was added at 1 μM, and incubation continued for another 10 minutes. Cells were then labeled with PE–annexin V and 7-AAD, and the RFI of the necrotic population was determined by FACS analysis as in Figure 2. The RFI at each concentration of competitive ligand was expressed as a percentage of the RFI of cells in its absence. (▪-▪), heparan sulfate; (▿-▿), unfractionated heparin; (▵-▵), 7 kd LMWH; (□-□), 6 kd LMWH; (●-●), 4.5 kd LMWH; (○-○), 3.5 kd LMWH; and (▴-▴), pentosan polysulfate.

This Feature Is Available To Subscribers Only