P-Rex and Vav family Rac-GEFs cooperate in peritoneal neutrophil recruitment. (A) Stronger impairment of neutrophil recruitment in P1V1 or P1V3 than P-Rex null or Vav null mice. Sterile peritonitis was induced with TGC, comparing GEF-deficient strains to wild-type mice of appropriate genetic background, and peritoneal lavages were performed after 1.5 hours, or 4.5 hours where indicated, and were analyzed for neutrophils or total leukocytes (right). Data are mean ± standard error of the mean (SEM) of 3 experiments with 5 to 16 mice/group; statistics Kruskal-Wallis test with Dunn’s multiple comparisons or Mann-Whitney U test between genotypes. (B) Time course. Sterile peritonitis was induced as mentioned earlier, except for different times. Data are mean ± SEM of n = 3 to 20 mice/time point and genotype; statistics 2-way analysis of variance (ANOVA) with Bonferroni multiple comparisons. (C) Normal vascular permeability. Evans Blue dye was IV injected 30 minutes before TGC or mock challenge, and peritoneal vascular permeability was assessed 1.5 hours after challenge. Data are mean ± SEM of 3 experiments with 5 to 23 mice/group; statistics unpaired Student t test between sham- and TGC-treated mice and 1-way ANOVA for genotypes. (D) Peripheral neutrophils and neutrophil mobilization. Mice were IV injected with 50 nM KC or were sham treated and neutrophil numbers in the circulation assessed after 1 hour. Data are mean ± SEM of n = 4 to 11 mice/genotype; statistics as in (B). (E) Bone marrow transplants. Mice were irradiated and their hematopoietic systems were reconstituted with donor bone marrow, as indicated, before TGC peritonitis was induced and neutrophil recruitment was determined, as in (A). Data are mean ± SEM of 3 experiments with 5 to 12 mice/group for P1V1 and with 4 to 11 mice/group for P1V3; statistics 1-way ANOVA.