Platelet Activation Marker Expression

The expression of platelet surface markers CD62p and PAC-1 were examined using flow cytometry (BD, FACSCanto plus, USA). The protocol has been previously described by Morel et al. [21 (link)]. Briefly, a part of whole blood samples were activated by ADP (20 µM) at 37 °C for 10 min. Resting and activated samples were fixed in 1% paraformaldehyde (PFA) at 37 °C for 1 h and stained with the corresponding antibodies: anti-CD61/PerCP, anti-CD62p/PE and anti-PAC-1/FITC (BD,San Diego, CA, USA) in dark at room temperature for 30 min. Then, samples were diluted in PBS until further use. Flow cytometry analysis was performed on 10,000 platelets (CD61/PerCP-positive). The results were presented as percentages of CD62p- and PAC-1-positive platelets in the samples.