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Fig. 6 | Cell Communication and Signaling

Fig. 6

From: cAMP- and cGMP-elevating agents inhibit GPIbα-mediated aggregation but not GPIbα-stimulated Syk activation in human platelets

Fig. 6

EB-induced Syk activation is not dependent on integrin αIIbβ3 outside-in signaling in contrast to the overall platelet aggregation. a Washed platelets (WP) from a patient with Glanzmann thrombasthenia (GT) and from a healthy donor (day control) were pre-incubated at 37 °C for 3 min in the presence or absence of 2 nM iloprost prior to stimulation with EB. Platelet aggregation was monitored until 60 s using light transmission aggregometry under stirring conditions and then stopped using Laemmli buffer for immunoblotting. b Platelet aggregation was stopped after 60 s using Laemmli buffer and Syk Y525/526 and Y352 were analyzed by immunoblotting. WP from a healthy donor were pre-incubated with iloprost (2 nM, 3 min) in the presence or absence of tirofiban (1.25 μg/ml, 1 min) prior to stimulation with EB. Platelet aggregation was stopped after 60 s using Laemmli buffer. Phosphorylation of c Syk Y525/526, e Syk Y352 and g PLCγ2 Y759 were analyzed by western blot. Quantification of p-Syk d Y525/526 and f Y352 compared to total Syk and h PLCγ2 Y759 compared to total PLCγ2. Data are from 3 different experiments with platelets from 3 healthy volunteers and presented as means ± S.D. ns: not significant, *p < 0.05

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