Skap2 regulates TNF-mediated integrin activation and outside-in signaling as well as neutrophil functions. (A) Soluble ICAM-1 binding of TNF-stimulated WT and Skap2^−/− neutrophils. n = 3. (B) Oxidative burst of WT and Skap2^−/− neutrophils stimulated with TNF in solution. n = 3. (C and D) Surface expression of CD11b (C) and CD18 (D) of TNF-stimulated WT or Skap2^−/− neutrophils. Quantification is shown below. n = 3. (E–G) Phagocytosis of pHrodo E. coli particles by WT and Skap2^−/− neutrophils. Phagocytosis (E), percentage of phagocytic cells (F), and representative images (G) of phagocytic neutrophils are shown. n = 3. (H) In vitro NET formation of WT and Skap2^−/− neutrophils stimulated with resting or TRAP-activated platelets. n = 3. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ^#, P < 0.05 versus all other time points; Student's t test. Data are means ± SEM. Ctrl, control.