PSGL-1 and CXCR2 signaling in monocytes does not enhance monocyte recruitment and tissue factor expression in deep vein thrombi. (A) Number of rolling M-CSFR (CD115)⁺ monocytes per minute per microscopic field in the IVC 3 hours after ligation (vertical axis). Circulating monocyte count for each genotype (horizontal axis). Inset, Representative image of adherent M-CSFR⁺ monocytes (red) in WT mice obtained with spinning-disk intravital microscopy. PE-conjugated anti–M-CSFR mAb was injected IV into WT mice 1 hour before surgery. Bar, 10 μm. (B) Quantification of endothelial surface area covered with firmly adherent monocytes. The data in the graphs of panels A and B represent the mean ± SEM from 5 mice in each group. (C) Number of M-CSFR⁺ monocytes per thrombus 24 hours after ligation, as measured by flow cytometry. (D) Normalized thrombus weight per M-CSFR⁺ monocyte. The data in panels C and D represent the mean ± SEM from 5 to 10 mice in each group. (E) Western blot of thrombus lysates probed with antibodies to fibrin, M-CSFR, and tissue factor. The Aα, Bβ, and γ chains of fibrin are marked. The data are representative of 3 experiments. (F) Normalized densitometric ratio of fibrin Aα chain to M-CSFR. (G) Normalized densitometric ratio of tissue factor to M-CSFR. The data in panels F and G represent the mean ± SD from 3 mice in each experimental group. *P < .05.