Impaired phagocytic activity and intracellular signaling but normal migratory ability of Hck ^−/− Fgr ^−/− Lyn ^−/− neutrophils. (A–C) WT or Hck^−/−Fgr^−/−Lyn^−/− neutrophils were stimulated with 1 mg/ml MSU crystals followed by assessment of phagocytosis of the MSU crystals (A and B), intracellular phosphorylation (C). Where indicated, the cells were pretreated with 10 μM cytochalasin D. (D and E) Mixed bone marrow chimeras carrying CD45.1-expressing WT and CD45.2-expressing WT, Itgb2^−/−, or Hck^−/−Fgr^−/−Lyn^−/− hematopoietic cells were subjected to MSU crystal–induced arthritis as described above. 24 h later, the hind paws were flushed, the ratio of CD45.1- and CD45.2-expressing neutrophils in the paw infiltrate was determined by flow cytometry and compared to the ratio in the peripheral blood. Panel D shows the results for each individual mouse, whereas panel E shows the accumulation of the CD45.2-expressing WT, Itgb2^−/−, and Hck^−/−Fgr^−/−Lyn^−/− neutrophils relative to the CD45.1-expressing WT cells. Panels A and C show representative images from two to three independent experiments. Panel B shows mean and SEM from two to six independent experiments. Panel E shows data mean and SEM of five to six chimeras per group from three independent experiments. Two-way ANOVA inhibitor × stimulus or genotype × stimulus interaction analysis (B) and one-way ANOVA (E); n.s., not significant; **, P < 0.01; ***, P < 0.001. See the text for actual P values. Source data are available for this figure: SourceData F4.