Myeloid Src-family kinases are required for MSU crystal–induced neutrophil activation. (A–I) WT or Hck^−/−Fgr^−/−Lyn^−/− neutrophils (A–I), along with various single and double knockout neutrophils (C) were stimulated with 1 mg/ml (A and C–I) or the indicated concentrations (B) of MSU crystals. (A–C) Luminometric measurement of reactive oxygen production. (D–I) Cells were incubated for 18 h followed by determination of cytokine, chemokine, and lipid mediator levels from cell-free supernatants using a commercial cytokine array (D) or ELISA assays (G–I). (E) Map of the different analytes on the cytokine array. (F) Quantification of cytokine array results by densitometry. Panels A (mean and SD) and D show results from a representative experiment. Bar graph shows mean and SEM from 4 (B), 3–8 (C), 2 (F), and 5–8 (G–I) independent experiments. Panels C and F show data normalized to WT MSU and WT control samples, respectively. Panel F shows analytes with at least fivefold MSU-induced increase in WT samples. RLU, relative luminescence unit; AUC, area under the curve. Two-way ANOVA genotype × stimulus interaction analysis; *, P < 0.05; **, P < 0.01; ***, P < 0.001. See the text for actual P values. Source data are available for this figure: SourceData F1.