OxLDL stimulates the activation of Syk and PLCγ2, leading to the phosphorylation of MLC. (A) Platelets (7 × 108) were treated with JNK inhibitor 1 (10 µM), PP2 (20 µM), or PP3 (20 µM) for 20 minutes followed by stimulation with oxLDL (50 µg/mL) for 15 seconds. Platelets were then lysed and Syk immunoprecipitated. Samples were subsequently separated by SDS-PAGE and were immunoblotted for phosphotyrosine, followed by reprobing for total Syk. (Ai) Representative blots. (Aii) Densitometric analysis of 3 independent experiments. *P < .05. (B) Platelets (7 × 108) were treated with PP2 (20 µM) or PP3 (20 µM) for 20 minutes, followed by stimulation with oxLDL (50 µg/mL) for 15 seconds. Platelets were then lysed and PLCγ2 immunoprecipitated. Samples were subsequently separated by SDS-PAGE and were immunoblotted for phosphotyrosine, followed by reprobing for total PLCγ2. (Bi) Representative blots. (Bii) Densitometric analysis of 3 independent experiments. *P < .05. All experiments were carried out in the presence of apyrase (2 U/mL), indomethacin (10 µM), and EGTA (1 mM). Densitometric data are expressed as mean ±SEM.
