Figure 5
Glycosaminoglycans are involved in RANTES-induced biological effects. (a,b) EPC were incubated with RANTES or [44AANA47]-RANTES or [E66A]-RANTES mutants (each at 3 nM). (a) EPC migration was assessed by Transwell assay. Results are expressed as mean ± SEM of EPC counted by field for three independent experiments. *P < 0.05 or ***P < 0.001 versus untreated cells (UT). § P < 0.05 versus RANTES. (b) EPC vascular sprout length was assessed in a 2D angiogenesis assay. Histogram represents cells treated with RANTES or [44AANA47]-RANTES or [E66A]-RANTES (each at 3 nM). *P < 0.05 versus untreated cells (UT). ££ P < 0.01 versus RANTES. (c) EPC migration induced by RANTES in absence of heparin was arbitrary set to 100%. The results of cells stimulated by 3 nM RANTES preincubated with a range of heparin concentrations (at 0.1, 1, 10 µg/mL) were expressed as a percentage of RANTES alone. *P < 0.05 versus cells stimulated by RANTES in the absence of heparin. (d) RANTES (3 nM) induced migration of EPC pre-treated with beta-D-xyloside (at 0.5 or 1 mM). The migration of untreated EPC towards RANTES was arbitrary set to100%. The results of cells treated with beta-D-xyloside and chemoattracted by RANTES were expressed as a percentage of untreated cells chemoattracted by RANTES. ***P < 0.001 versus untreated cells (UT) chemoattracted by RANTES. (e,f) EPC area induced by RANTES (3 nM) was assessed after a cell spreading assay (e) and the length of vascular sprouts formed by EPC was assessed after a 2D angiogenesis assay (f). *P < 0.05, cells stimulated by RANTES pre-incubated with 10 µg/mL heparin versus cells stimulated by RANTES in absence of a preincubation with heparin. & P < 0.05 cells treated with 1 mM beta-D-xyloside and chemoattracted by RANTES versus untreated cells chemoattracted by RANTES.
