Skap2 regulates TNF-mediated neutrophil recruitment. (A–C) IVM of TNF-inflamed postcapillary venules of WT and Skap2^−/− mice. (A) Cumulative histogram of rolling velocities. (Inset) Data in a bar graph. 175 data points for each genotype are shown. (B and C) Number of adherent cells (B) and number of extravasated cells (C) 2 h after TNF application. n = 4 mice/group. (D) Representative images of transmigrated cells 2 h after TNF application. (E–G) IVM of TNF-inflamed postcapillary venules of WT mice after injection of differentially ex vivo–labeled WT and Skap2^−/− bone marrow cells. Rolling velocities (E), number of adherent cells (F), and number of extravasated cells (G) 2 h after TNF application are shown. The percentage of total fluorescent cells for each genotype is shown. n = 3 mice/group. (H) Peripheral blood cell counts of WT (n = 7) and Skap2^−/− (n = 5) mice. (I and J) Expression of the indicated myeloid maturation markers and cell surface receptors on WT and Skap2^−/− neutrophils. Black lines show WT, gray dashed lines show Skap2^−/−, and black dotted lines show isotype control. Quantification is shown below. n = 3. *, P < 0.05; **, P < 0.01; Student's t test. Data are means ± SEM.