Reelin enhances monocyte–endothelial cell adhesion and blunts VEGF-induced eNOS activation through Apoer2. Reelin enhances monocyte–endothelial cell adhesion.

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Reelin enhances monocyte–endothelial cell adhesion and blunts VEGF-induced eNOS activation through Apoer2. Reelin enhances monocyte–endothelial cell adhesion and blunts VEGF-induced eNOS activation through Apoer2. (A) Representative images of U937 monocytes (small, round cells) adhered to HAECs (cobblestone shape) treated with vehicle, mock, or Reelin. Scale bar, 200 μm. (B to F) Summary plots depicting means ± SEM of the following: (B) number of adherent monocytes per 40× field of view in (A) (n = 6 independent experiments; ****P < 0.0001). (C) Monocyte-HAEC adhesion with mock, Reelin ± mouse immunoglobulin G (IgG), or CR50 (n = 6 independent experiments; ***P < 0.002, ****P < 0.0001). (D) Monocyte-HAEC adhesion with vehicle, mock, or Reelin (top) after transfection with either control [control RNA interference (RNAi)], double-stranded RNA targeting Apoer2 (left; n = 3 independent experiments; ****P < 0.0001), or Vldlr (right; n = 3 independent experiments; ****P < 0.0009). In parallel, whole-cell lysates were immunoblotted for Vldlr or Apoer2 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (bottom). (E) VEGF-stimulated eNOS activity in HAECs with or without Apoer2 knockdown, pretreated with or without mock or Reelin (n = 6 independent experiments; *P < 0.02, **P < 0.006, ****P < 0.0001). (F) Monocyte-HAEC adhesion in the presence of mock or Reelin ± SNAP (n = 6 independent experiments; **P = 0.0081, ***P = 0.0001). Additional statistical details are summarized in table S1. Yinyuan Ding et al., Sci. Signal. 2016;9:ra29 ©2016 by American Association for the Advancement of Science