Skap2 regulates integrin-mediated outside-in signaling. (A–D) Neutrophils from WT or Skap2^−/− mice were plated on pRGD and adhesion (A) and spreading (B and C) were analyzed. (D) Representative images of spread neutrophils. 75 cells/experiment were analyzed. n = 3. (E–G) Adhesion-dependent oxidative burst of WT and Skap2^−/− neutrophils plated on pRGD alone (E) or ICAM-1 (F) and fibrinogen (G) in the presence of TNF. n = 4. (H) Oxidative burst of WT and Skap2^−/− neutrophils stimulated with PMA in solution. n = 4. (I and J) WT and Skap2^−/− neutrophils were plated on pRGD for 10 min, and lysates were immunoblotted with anti–p-ERK1/2 and anti-ERK1/2 (I) or anti–p-Akt and anti-Akt (J) and anti–α-tubulin antibody. Quantification is shown on the right. n = 4. *, P < 0.05; **, P < 0.01; ^#, P < 0.05 versus all other time points; Student's t test. Data are means ± SEM. Ctrl, control.