Figure 2

In vitro and in vivo functional effects of 3F12E7 anti-FGF2 scFv. 3F12E7 scFv reduces in vitro endothelial cell proliferation (a) and migration (b). Cells were incubated with 10 µg/mL of the indicated mAbs. No difference was detected between 3F12E7 scFv and 3F12E7 full-length IgG groups. Cell proliferation and migration were accessed by trypan blue exclusion and scratch assays, respectively. Representative micrographs of the scratch assay are on (c). Dashed lines indicate original wound edges. Scale bar, 200 µm. *P < 0.05 compared to isotype and vehicle controls; one-way ANOVA/Bonferroni’s post-test. (d) Immunoblotting analyses of ERK1/2 phosphorylation in HUVEC after 48-h incubation with the indicated mAbs (50 µg/mL). β-actin was used as loading control. Graph shows the quantitative densitometry of the immunoblot results. Data are mean ± s.d. of the relative intensity of the bands, normalized to that of isotype ctrl IgG group, from two independent assays. The full-length image scans and the result of an additional independent assay are provided in Supplementary Fig. S4a. (e, f) 3F12E7 scFv reduces xenograft tumor growth similarly to 3F12E7 full-length IgG mAb. (e) Tumor growth curve. (f) Excised tumor mass on day 12. Treatment started four days after subcutaneous injection of B16-F10 cells. Result (mean ± s.d.) is representative of two independent experiments. Experimental groups: isotype control full-length IgG antibody, n = 6; 3F12E7 full-length IgG mAb, n = 6; 3F12E7 scFv, n = 6. *P < 0.05 compared with isotype control group; one-way ANOVA/Bonferroni’s post-test.