Figure 7
MMRN2495–674 inhibits angiogenesis in vitro. (a) Representative flow cytometry histograms of CLEC14A-ECD-Fc pre-incubated with MMRN2495–674 or MMRN2495–603 and then used to stain HUVEC surface, hFc used as isotype control. (b) MMRN2495–674 significantly blocked cell surface binding. (*P<0.05 Mann–Whitney test n=4). (c) Representative images of MMRN2495–603 and MMRN2495–674 coated plates with adherent HUVEC stained with crystal violet; scale bar, 150 μm. (d) HUVEC adhered significantly more to MMRN2495–674 compared with MMRN2495–603 or bovine serum albumin (BSA; **P<0.01 Mann–Whitney test, n=6). (e) HUVEC were added to Matrigel and treated with PBS, hFc, CLEC14A-ECD-Fc (CLEC14A-Fc) MMRN2495–603 or MMRN2495–674. Representative images from 6, 12 and 24 h post Matrigel plating from one of three independent experiments; scale bar, 100 μm. (f) Quantification of number of meshes at 6, 12 and 24 h time points, average values from eight fields of view from three independent experiments, (*P<0.05, **P<0.01 unpaired t-test, n=3, error bars represent s.e.m.). (g) Representative images of HUVEC-fibroblast co-culture assay from three different umbilical cords. Scale bar, 800 μm. (h) Quantification of number of tubules and total tubule length, from averages of four fields of view from three independent experiments. (*P<0.05 unpaired t-test, n=3, error bars represent s.e.m.).
