Total plcγ2

Human thrombin, PGE1, dimethyl sulfoxide (DMSO), ADP, fibrinogen, TXA₂ analog U46619, ferric chloride (FeCl₃), and all the reagents were purchased from Sigma (St. Louis, MO, United States). D-Phe-Pro-Arg-chloromethyl ketone (PPACK) was purchased from EMD Millipore (Billerica, MA, United States). Collagen-related peptide (CRP) was obtained from Dr. Richard Farndale (Department of Biochemistry, University of Cambridge, United Kingdom). Phycoerythrin (PE)-conjugated isotype control IgGs, rat monoclonal antibodies against mouse P-selectin, and activated αIIbβ3 (JON/A) were obtained from Emfret Analytics (Eibelstadt, Germany). Antibodies against phospho-PLCγ2 at Tyr759, phospho-PLCβ3 at Ser1105, phospho-Akt at Ser473, Total Akt, Total PLCγ2, Total PLCβ3, and actin were obtained from Cell Signaling (Danvers, MA, United States). Calcium dye (FLIPR Calcium Assay kit) was obtained from Molecular Devices (Sunnyvale, CA, United States).

Human thrombin, PGE1, rhodamine-phalloidin, dimethyl sulfoxide (DMSO), ADP, fibrinogen, human fibrinogen, ferric chloride (FeCl₃), Acetylsalicylic acid (ASA), carboxymethylcellulose (CMC), and all the reagents were purchased from Sigma (St. Louis, MO, USA). Equine tendon collagen (type I) and ATP luciferin/luciferase reagent were obtained from Chrono-log (Havertown, PA, USA). CRP was obtained from Dr Richard Farndale (Department of Biochemistry, University of Cambridge, UK). Phycoerythrin (PE)-conjugated isotype control IgGs, rat monoclonal antibodies against mouse P-selectin, activated αIIbβ3 (JON/A) were from Emfret Analytics (Eibelstadt, Germany). Antibodies against phospho-Syk at Tyr525/526 (Cat# 2710S), phospho-PLCγ2 at Tyr1217 (Cat# 3871S), phospho-PI3K p85α/β at Tyr458/p55α/γ at Tyr199 (Cat# 4228S), phospho-Akt at Ser473 (Cat# 9271S), phospho-ERK at Thr202/Tyr204 (Cat# 4337S), Total Syk (Cat# 12358S), Total PLCγ2 (Cat# 3872S), Total PI3K p85 (Cat# 4249S), Total Akt (Cat# 9272S), Total ERK (Cat# 9102S), and β-actin (Cat# 4967S) were obtained from Cell Signaling (Danvers, MA, USA). Monoclonal antibodies against phospho-integrin β3 at Tyr 759 and Total integrin β3 were obtained from Santa Cruz (Santa Cruz, CA, USA). Calcium dye (FLIPR Calcium Assay kit) was from Molecular Devices (Sunnyvale, CA, USA).