Figure 5.

RMP migration is mediated through CXCR4, but CXCR7 is required for transendothelial migration and mediates EC adhesion. (A) Dose-dependent migration of RMPs in response to SDF-1. Results are expressed as the mean ± the SEM of triplicate assessment. One representative of three independent experiments is shown. *, P < 0.05; **, P < 0.001. (B) RMP migration is reverted by pretreatment with anti-CXCR4 neutralizing antibody, but not by an anti-CXCR7 neutralizing antibody. Results are expressed as the mean ± the SEM as assessed in four independent experiments. **, P < 0.001. (C) Transendothelial migration of RMPs is reverted by pretreatment with either an anti-CXCR4 or an anti-CXCR7 neutralizing antibody. One representative of three independent experiments is shown. (D) Quantitation of transendothelial migration of RMPs. Results are expressed as the mean ± the SEM of fluorescence intensity as observed in four independent experiments. a vs. b and b vs. c: P < 0.05; a vs. c: NS. (E) Quantitation of transendothelial migration of RMPs. Results are expressed as the mean ± the SEM of fluorescence intensity as observed in four independent experiments. a vs. b and b vs. c: P < 0.05; a vs. c: NS. (F) HUVECs were stained with Ulex europeus I lectin (green) to visualize the EC monolayer. To-Pro-3 was used to counterstain nuclei. (G) PKH26-labeled RMPs (red) were cultured in presence or absence of an isotype control antibody, an anti-CXCR7 antibody or an anti-CXCR4 neutralizing antibody. One representative of three independent experiments is shown. (H) Quantitation of RMP adhesion to HUVEC monolayers. Results are expressed as the mean ± the SEM of fluorescence intensity as observed in four independent experiments. **, P < 0.001. Bars; (C and F) 50 μm; (G) 100 μm.