Figure 6

Effect of clec14a-CTLD IgGs on endothelial cell proliferation and activation. (a) HUVECs were incubated in the absence (MOCK) or presence of clec14a-CTLD IgGs, cetuximab or 5-FU (positive control) for 2 days. Cell viability was assessed by measuring absorbance at 450 nm. Values represent mean±s.d. of triplicate measurements from one of two independent experiments. (b) HUVECs cultured in the absence or presence of clec14a-CTLD IgG (clone 1) were stained with rhodamine–phalloidin and examined by confocal microscopy. (c) COS-7 cells transfected with GFP or clec14a-GFP were incubated in the absence or presence of clec14a-CTLD IgG (clone 1). Cell viability was assessed by measuring absorbance at 450 nm. Values represent the mean±s.d. of triplicate measurements from one of two independent experiments. (d) HUVECs were cultured in the absence (dashed line) or presence (solid line) of hTNFα, clec14a-CTLD IgGs or cetuximab; stained with anti-VCAM-1 (upper) or ICAM-1 (lower) polyclonal antibody and analyzed by flow cytometry. hTNFα served as a positive control for endothelial cell activation. Results are representative of three independent experiments.