Blocking PI3Kγ in Plcg2^−/− neutrophils completely abolishes E-selectin–mediated slow rolling. (A) Rolling velocity of WT neutrophils on E-selectin alone or E-selectin/ICAM-1 of WT mice pretreated with either a PI3Kδ-inhibitor (PI3Kδ-inh.) or DMSO. (B) Rolling velocity of WT and Plcg2^−/− neutrophils on E-selectin alone or E-selectin/ICAM-1 of either PI3Kγ-inhibitor (PI3Kγ inh.)– or DMSO-pretreated mice. (C) Rolling velocity of WT and Pik3cg^−/− neutrophils on E-selectin or E-selectin/ICAM-1 of either untreated or PLC-inhibitor–pretreated mice. Data are presented as means ± SEM from 3 mice. (D) Mixed chimeric mice were generated by injecting bone marrow cells from Pik3cg^−/− mice and LysM-GFP⁺ WT mice into lethally irradiated WT mice. Cumulative histogram of rolling velocities of 100 GFP⁺ (WT; ●) and 100 GFP⁻ (Pik3cg^−/−; ○) leukocytes in inflamed cremaster muscle venules of mixed chimeric mice (n = 3) treated with PTx and a monoclonal blocking P-selectin antibody (RB40.34). Inset data are means ± SEM. (E) Numbers of adherent cells per square millimeter in murine cremaster muscle venules. The cremaster muscle was exteriorized 2 hours after intrascrotal injection of 500 ng TNF-α in chimeric mice reconstituted with bone marrow from Pik3cg^−/− mice or WT mice. The dotted line indicates the number of adherent cells in WT mice treated with PTx and monoclonal blocking E-selectin antibody (9A9). #P < .05.