Figure 1

Del-1 is associated with glycocalyx and is detached from the cell surface by heparinase or PI-PLC treatment. (A,B) Binding of Del-1 (50 nM) to immobilized BSA, HS (50 nM), and SDC1 (50 nM) and binding of Del-1 to immobilized HS or SDC1 upon heparinase treatment (0.005 U/ml or 0.05 U/ml), as assessed by solid-phase binding assay. The data are shown as the mean ± SD (n = 3 mice per group). ^*P < 0.05 and ^†P < 0.01, Student’s t-test. (C) Immunohistochemistry for Del-1 treated with heparinase (0.05 U/ml) or PI-PLC (2 U/ml) at 37 °C for 1.5 h. Nuclei were stained with DAPI (blue). Scale bars, 50 μm. (D) ELISA for Del-1 from the supernatants from the culture treated with heparinase (0.5, 2 U/ml) or PI-PLC (0.2, 2 U/ml) at 37 °C for 1 h. The data are shown as the mean ± SD (n = 6 mice per group). ^*P < 0.05 and ^†P < 0.01, Student’s t-test. BSA: bovine serum albumin; DAPI: 4′,6-diamidino-2-phenylindole; Del-1: developmental endothelial locus-1; ELISA: enzyme-linked immunosorbent assay; HS: heparan sulfate; PI-PLC: phosphatidylinositol-specific phospholipase C; SDC1: syndecan-1.