Fig. 6.
Fig. 6. Effect of intracellular cAMP and cGMP modulation on GPIbα-mediated [Ca++]i elevations in platelets interacting with immobilized VWF. / A blood cell suspension (see the legend for Figure 2) was supplemented with the anti-αIIbβ3 antibody LJ-CP8 (100 μg/mL) and 2 mM EGTA to block [Ca++]ielevations not mediated by GPIbα. Perfusion and analysis were performed as described in the legend for Figure 5. Before perfusion, samples were incubated at 37°C with either theophylline for 15 minutes (A), sildenafil for 20 minutes (B), or dibutyryl cAMP or 8Br-cGMP for 10 minutes (C). Theophylline was used at a high concentration to offset adsorption by erythrocytes.22 The number of platelets with at least one [Ca++]ielevation of type α/β (no γ peaks were observed in these experiments) was measured, and results are shown normalized to the values observed with control blood cell suspensions left untreated. The data represent the mean ± 95% CI from 5 different experiments.

Effect of intracellular cAMP and cGMP modulation on GPIbα-mediated [Ca++]i elevations in platelets interacting with immobilized VWF.

A blood cell suspension (see the legend for Figure 2) was supplemented with the anti-αIIbβ3 antibody LJ-CP8 (100 μg/mL) and 2 mM EGTA to block [Ca++]ielevations not mediated by GPIbα. Perfusion and analysis were performed as described in the legend for Figure 5. Before perfusion, samples were incubated at 37°C with either theophylline for 15 minutes (A), sildenafil for 20 minutes (B), or dibutyryl cAMP or 8Br-cGMP for 10 minutes (C). Theophylline was used at a high concentration to offset adsorption by erythrocytes.22 The number of platelets with at least one [Ca++]ielevation of type α/β (no γ peaks were observed in these experiments) was measured, and results are shown normalized to the values observed with control blood cell suspensions left untreated. The data represent the mean ± 95% CI from 5 different experiments.

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