The p110γ is required for efficient capture of neutrophils by E-selectin. (A) Immunoprecipitates of spleens from WT mice using a p110δ-specific antibody that was assayed for PIP3 production in the absence (▪) or presence (▪)of IC87114 (10 μM). The results are expressed as the percent activity in vehicle-treated p110δ immunoprecipitates and represent the mean plus or minus SD values of 3 independent determinations in duplicate. *P < .01 as compared with vehicle-treated WT spleen lysate. Rolling fraction of P-selectin–/–/GFP–/+ animals (B) pretreated with vehicle control (▪), IC87114 (▪), or mAb 9A9 (▦) 1 hour prior to induction of inflammation with TNFα or (C) in those that also lacked p110γ. Data represent the mean plus or minus SD and are normalized as a percentage of control; *P < .05 as compared with control. n = number of mice /venules analyzed. (D) In vivo imaging of E-selectin expression on CM venules in mice with or without prior stimulation with TNFα. Fluorescent Qdots coated with an antibody that recognizes murine E-selectin were injected intravenously into animals and immunolocalization of this adhesion molecule was visualized by intravital microscopy. Transmitted light (i) and epifluorescence (ii) images depict staining of venules in P-selectin–/– mice in the absence of TNFα stimulation. Representative micrographs of inflamed venules in P-selectin–/– animals pretreated with (iii) IC87114 or (iv) lacking p110γ, respectively, or mice deficient in E-selectin in the absence (v) or presence (vi) of IC87114.
