Skip to main content
. 2014 Oct 14;3(5):e001369. doi: 10.1161/JAHA.114.001369

Figure 5.

Figure 5.

Endothelial cell repair inhibition by antiphospholipid antibodies (aPLs) requires β2‐glycoprotein I (β2GPI) and apolipoprotein E receptor 2 (ApoER2). A, Scratch assays were performed with bovine aortic endothelial cells (BAECs) treated with media alone (basal), vascular endothelial growth factor (VEGF; 50 ng/mL), VEGF+isotype‐matched control IgG (BBG, 0.3 μg/mL), or VEGF+3F8 monoclonal antibody to β2GPI (0.3 μg/mL). Values are mean±SEM, n=8/group, *P<0.05 vs basal, † P<0.05 vs BBG. B, BAECs were transfected with control silent interfering RNA (siRNA) or siRNA targeting ApoER2, and whole‐cell lysates were immunoblotted for ApoER2 and actin. C, Scratch assays were performed 48 hours later with treatments with media alone (basal), VEGF (50 ng/mL), VEGF+human IgG from healthy nonautoimmune individuals (NHIgG; 100 μg/mL), or VEGF+aPLs (100 μg/mL) for 24 hours. Values are mean±SEM, n=4 for no treatment (ApoER2 siRNA) and VEGF, n=6 for no treatment (Control siRNA), VEGF+NHIgG, and VEGF+aPLs*P<0.05 vs basal, †P<0.05 vs NHIgG, #P<0.05 vs Control siRNA. D, Scratch assays were performed 48 hours after cell transfection with control siRNA or siRNA targeting ApoER2, using the treatments described for A. Values are mean±SEM, n=4 for no treatment and VEGF treatment group, and n=8 for VEGF+BBG and VEGF+3F8 treatment group, *P<0.05 vs basal, †P<0.05 vs BBG, #P<0.05 vs Control siRNA. E, Male ApoER2+/+ or ApoER2−/− littermates treated with NHIgG (100 μg/mouse) or aPLs (100 μg/mouse) underwent carotid artery endothelial denudation, and the remaining area of denudation was evaluated 72 hours later. Representative images of the carotid artery intimal surface displaying Evans blue dye incorporation are shown. F, Summary data for the area of remaining denudation at 72 hours are expressed in arbitrary units. Values are mean±SEM. For ApoER2+/+ n=4 (uninjured, median was 161.5) and n=5 (injured, median was 19 087). For ApoER2−/−, n=5 (uninjured, median was 7736) and n=5 (injured, median was 5923). *P<0.05 vs NHIgG in ApoER2+/+, †P<0.05 vs aPLs in ApoER2+/+.