Figure 5:

P-selectin-PSGL1 interactions mediate adult platelet induced trafficking monocyte phenotype. Adult or P7 platelets were co-incubated with monocytes with IgG or anti-PSGL1 Ab. 4 hrs later PMAs, Ly6C, CCR2 and CCR5 expression were determined. A) Adult platelet associated PMAs were reduced and B) Ly6C was unchanged by anti-PSGL1 Ab. C-D) Adult platelets increased CCR2 and CCR5 compared to P7 platelets, which was decreased by anti-PSGL1 antibody. E-H) Monocyte trafficking in vitro. Adult or P7 platelets and monocytes were co-cultured in the top of a transwell chamber with control IgG or anti-PSGL1. CCL2 or CCL5/RANTES were placed in the bottom chamber and monocyte migration determined. E-F) Adult platelets induced monocyte trafficking to CCL2 which was decreased by anti-PSGL1 Ab. E) Representative Image and F) Quantification. G-H) Adult platelets induced monocyte trafficking to CCL5, which was reduced by anti-PSGL1 Ab. G) Representative Image and H) Quantification (mean ± SEM, 1-way ANOVA with Bonferroni correction). Scale bar =200 μm.