Fig. 1.

Echicetin-coated polystyrene beads activate Syk tyrosine kinase and the PI3K-dependent serine/threonine kinase Akt. a Human washed platelets (WP) were stimulated under stirring conditions with EB . Platelet aggregation (see Fig. S1C, D) was stopped after 1, 2 or 5 min by adding Laemmli buffer. Tyrosine phosphorylation of Syk, LAT, PLCγ2 and serine phosphorylation of Akt were analyzed by immunoblotting compared to the total protein respectively. b The kinetics of the phosphorylation patterns represent means ± S.D. of 3 independent experiments with platelets from 3 healthy donors. c WP were pre-incubated for 5 min at 37 °C with vehicle control (DMSO) or with the PI3K inhibitor wortmannin (100 nM final concentration) prior to stimulation with EB. d Quantitative data of Akt S473 phosphorylation compared to the loading control α-actinin are represented as means ± S.D. of 3 independent experiments with platelets from 3 healthy donors (Samples were loaded on the same gel; a black line was used to indicate that a group of samples not related to this data set was not shown); ns: not significant, ****p < 0.0001