Fig. 2.

Analysis of platelet kinase signaling pathways. Platelets were either pre-treated with DMSO (−) or PF-228 (+) (1 μM) for 5 min before stimulation with CRP (5 μg/ml) for 3 min under stirred conditions before lysis; basal samples (−) were lysed after 8 min. FAK was immunoprecipitated from platelet lysates before immunoblotting with phosphotyrosine to assess activation (A). Lysates were also prepared by lysis of platelets after treatment in sample buffer. Aliquots (15 μl) from these lysates were resolved by SDS–PAGE and immunoblotted with the activation specific antibodies indicated in (B). Samples were also blotted with anti-actin to confirm equal extraction of protein from platelet samples. All blots are representative of three independent experiments.