Figure 2
Chemotactic response of mouse neutrophils to rhAng-1 (A) and rhAng-2 (B). Dose-response effects of rhAng-1 and rhAng-2 on isolated mouse peripheral blood neutrophil chemotaxis. KC (10−7 M) was used as a positive control. Results presented as mean ± SEM (n = 4) and analysed for statistical significance using ANOVA followed by Dunnett’s comparing all to buffer control. *P < 0.05 **P < 0.01 ***P < 0.001. The effect of isotype control (rat IgG1), anti-Tie2 (12 µg/ml; 6 µg/10−6 cells) (C) or anti-CD18 (GAME-46) (D) (15 µg/ml; 7.5 µg/10−6 cells) on chemotaxis induced by KC (10−7 M) or rhAng-1 (0.7 µg/ml). Results presented as mean ± SEM (n = 3–4) and analysed for statistical significance using two-way ANOVA followed by Bonferroni’s test for multiple comparisons. *P < 0.05 **P < 0.01 ***P < 0.001 compared to isotype control. (E) Neutrophils were treated with rhAng-1 (0.7 µg/106 neutrophils) or PBS and subjected to extracellular crosslinking. Protein levels of Ang-1 co-immunoprecipitated with CD18 were detected by immunoblotting and quantified using densitometry. Results are expressed as mean ± SEM, n = 3 and analysed for statistical significance using a paired t-test.
