Figure 6
Expression of G protein-coupled receptors and proteoglycans on EPC. (a) Flow cytometry analysis of cells incubated with anti-CCR1, anti-CCR5, anti-SDC-1, anti-SDC-4, anti-CD44, anti-heparan sulfate (HS) antibodies or their respective isotypes and with Alexa Fluor-488 labeled secondary antibodies. Data shown are representative of three independent experiments. (b) Inhibition of biotinylated RANTES (40 nM) binding by preincubation of EPC with anti-CCR5, anti-SDC-4, anti-CD44 antibodies or their respective isotypes. Binding of B-RANTES was analyzed by flow cytometer with avidin-FITC control condition. Reactivity was compared to avidin-FITC. (c) The effect of G protein-coupled receptors and proteoglycans in RANTES-mediated cell spreading (c), migration (d) and 2D angiogenesis assays (e, f) was assessed after preincubation of EPC with anti-CCR5, anti-SDC-4, anti-CD44 antibodies or their respective isotypes. RANTES-induced EPC spreading, migration, vascular sprout area and length of EPC pre-incubated with the isotype were arbitrary set to 100%. *P < 0.05 versus cells preincubated with specific isotypes.
