Gα_i-independent neutrophil recruitment is defective in Btk^−/−and Plcg2^−/− mice. (A) Number of extravasated leukocytes in cremasteric venules of TNF-α–treated chimeric mice reconstituted with bone marrow from WT mice (n = 4), Btk^−/− mice (n = 4), Pik3cg^−/− mice (n = 3), or Plcg2^−/− mice (n = 4) per 1.5 × 10⁴ μm² tissue area. The measurements were performed 2 hours after intrascrotal TNF-α injection. The same groups were also analyzed after pretreatment with 4 μg PTx intravenously (+PTx; WT mice + PTx [n = 4], Btk^−/− mice + PTx [n = 4], Pik3cg^−/− mice + PTx [n = 3], Plcg2^−/− mice + PTx [n = 4]). In addition to this, we analyzed WT mice after pretreatment with a blocking E-selectin antibody alone (+ anti–E-sel. ab [n = 3]) and in combination with PTx (WT mice + anti–E-sel. ab + PTx [n = 4]). (B) Neutrophil influx into the peritoneal cavity 8 hours after 1 mL injection of 3% thioglycollate into chimeric mice reconstituted with bone marrow from WT mice (n = 5), Btk^−/− mice (n = 5), or Plcg2^−/− mice (n = 5). The same groups were also analyzed after pretreatment with 4 μg PTx intravenously (+PTx; WT mice + PTx [n = 5], Btk^−/− mice + PTx [n = 4], and Plcg2^−/− mice + PTx [n = 5]). Total numbers of neutrophils in the peritoneal lavage fluid were determined by flow cytometry and hemocytometer count. #P < .05.