Statins modulate PECAM-1 tyrosine phosphorylation. Washed human platelets were incubated with simvastatin or control for 5 minutes under conditions that disfavor aggregation (presence of EGTA [1 mM], indomethacin [10 mM], apyrase [2 U mL-1]). PECAM-1 was immunoprecipitated from cell lysates using anti–PECAM-1 antibody (WM59) and immunoblotted with an anti-phosphotyrosine antibody (4G10). Blots are representative of 3 separate experiments and normalized to loading control. Numerical data represent the percentage increase above phosphorylation levels in nontreated platelets (A). Washed human platelets were treated with PECAM-1 antibody to mediate clustering (PECAM-1 XL, 0.8 μg mL-1) or isotype control (IgG-XL, 0.8 μg mL-1) prior to CRP-XL (1 μg mL-1) stimulation in the absence or presence of simvastatin (10 μM) and aggregation was measured (B). Washed platelets were incubated with PECAM-1 antibody to mediate clustering (PECAM-1 XL) or isotype control (IgG-XL) (1 μg mL-1) prior to CRP-XL (1 μg mL-1) stimulation in the absence or presence of simvastatin (5-10 μM). PECAM-1 was immunoprecipitated from cell lysates. Immunoprecipitates were divided in 2, with half probed for tyrosine phosphorylation and half probed for PECAM-1, to ensure equivalent levels of protein isolation between samples. Blots are representative of 3 separate experiments and normalized to loading control (C), t test, *P ≤ .05, ***P ≤ .001. A vertical line has been inserted to indicate the position where a gel lane was removed.