Antiplatelet agents and TKIs differentially reduce oxLDL potentiation of platelet GPVI/ITAM signaling. Replicate samples (n = 3) of washed human platelets (1 × 109/mL; in modified HEPES/Tyrode buffer without addition of calcium) were pretreated with P2YR inhibitor ticagrelor, BTK inhibitor ibrutinib (10 μM), or Syk inhibitor R406 (10 μM) for 10 minutes and then stimulated with oxLDL (20 μg/mL) in combination with GPVI-specific agonist CRP-XL (0.5 μg/mL) for 5 minutes at 37°C. After collection into sample buffer, platelet lysates were separated by capillary electrophoresis and stained with indicated antisera for quantitative immunoblot analysis on a ProteinSimple Jess western blotting system. (A) Representative immunoblot analysis and (B) quantitation of signal intensities with indicated antisera; α-tubulin serves as a loading control for total protein levels. Positions of molecular weight (kD) markers are indicated.
