Figure 1

Reelin induces clustering of EphB2 proteins. (A) Stimulation with preclustered ephrin B1 extracellular domain or Reelin induces clustering of EphB2 (green) in stably transfected NG-108 cells. (B) Clustering of endogenous EphB2 (red) in primary cortical neurons after stimulation with preclustered ephrin B1 or Reelin. Clustering is observed in wild-type (WT) neurons as well as in neurons lacking both ApoER2 and VLDLR (VR^−/−;ER2^−/−). Scale bar, 5 μm. (C) Quantification of EphB2 cluster density in ephrin B1- or Reelin-treated primary neurons prepared from WT or ApoER2/VLDLR double knockout embryos (VR^−/−;ER2^−/−) (S.E.M., ^***P < 0.001 compared with control, n = 15 per condition, 3 knockout embryous). (D) Quantification of EphB2 cluster density in ephrin B1- or Reelin-treated primary neurons preincubated with GST or GST-RAP, a lipoprotein receptor antagonist (S.E.M., ^***P < 0.001 compared with control, n = 15 per condition). (E) Co-localization of EphB2 and Reelin in cortical neurons. Cultured cortical neurons were treated with recombinant Reelin, washed twice, fixed and immunostained at DIV 6 for EphB2 and Reelin. Scale bar, 10 μm. A higher magnification of a dendrite is shown (boxed area). Co-localization of EphB2 (red) and Reelin (green) appears as yellow puncta.