Platelet Aggregation Assay by LTA

Aggregation of washed human platelets was monitored by light transmission aggregometry (LTA) using an Apact4S Plus aggregometer (DiaSys Greiner, Flacht, Germany). Platelets were stimulated by ADP (Sigma-Aldrich, Saint Louis, MO, USA), convulxin (Enzo life sciences, Lausen, Switzerland), echicetin beads (EB) (prepared as previously described [28 (link),53 (link)]), or PDBu (Sigma-Aldrich, Saint Louis, MO, USA) under stirring conditions (1000 s⁻¹) at 37 °C. Platelets were pre-treated with different effectors prior to stimulation: Src family kinase inhibitor PP2 (Abcam, Cambridge, UK), Syk inhibitors OXSI-2 (Merck, Darmstadt, Germany) and PRT-060318 (Sellckem, Houston, TX, USA), PKC pan-inhibitor GF109203X (Enzo life sciences, Lausen, Switzerland), or cAMP-elevating agent iloprost (Bayer AG, Leverkusen, Germany).