<p>(A, B) Total RNA was extracted from central marrow EC, endosteal marrow EC, spleen EC, central marrow SC and endosteal marrow SC. The mRNA expression level of Tie-2 and VE-Cadherin was measured using quantitative RT-PCR. The relative expression was normalized to Hypoxanthine-guanine phosphoribosyltransferase (HPRT) levels and calculated from standard curves. (*p<0.05, **p<0.01; n = 6 from 3 independent experiments; error bars represent standard deviation). (C) Cells cultured in endothelial or stromal culture conditions were stained with antibodies to CD31 and VE-Cadherin. Positive signals were visualized with FITC conjugated secondary antibody. Nuclei were visualized with 4,6-diamidino-2-phenylindole (DAPI). (D) Central marrow EC, endosteal marrow EC, spleen EC, central marrow SC and endosteal marrow SC were harvested by enzyme-free cell dissociation solution. The expression of CD31 and CD45 was analyzed by flow cytometry with PE conjugated anti-CD31 and APC-Cy7 conjugated anti-CD45. (E) Cells cultured in endothelial or stromal culture conditions were stained with alkaline phosphatase activity. Positive reaction was observed as dark blue violet in the cells.</p
