FIGURE 5.

Reelin internalization and degradation rates of ApoER2 and VLDLR depend on their sorting within the plasma membrane. 3T3 cells expressing ApoER2 or one of the chimeric receptors containing the extracellular domain of ApoER2 (A) or VLDLR or one of the chimeric receptors containing the extracellular domain of VLDLR (B) were incubated with RCM at 4 °C to allow binding of Reelin to the respective receptors. Cells were then shifted to 37 °C for the indicated time periods to allow internalization and degradation of the ligand. After washing the cells, extracts were prepared and analyzed for cell-associated Reelin by Western blotting using Ab G10 in combination with an HRP-coupled goat anti-mouse antibody. C, 3T3 cells expressing ApoER2 were treated as described for A in the presence (lower panel) or absence (upper panel) of the raft-disrupting agent CDX (5 mm). Cell extracts were analyzed as described for A.