Skip to main content
. 2010 Jan;24(1):58–69. doi: 10.1096/fj.09-138123

Figure 5.

Figure 5.

Reelin alters interaction between X11α and ApoEr2, and X11α increases cell-surface ApoEr2 in MCF 10A cells. A) Cell lysates from COS7 and MCF 10A cells were Western blotted for APP, ApoEr2, Dab1, X11α, and FE65 to demonstrate expression. COS7 cells expressed detectable levels of APP and ApoEr2, but no expression of Dab1 and X11α with a high level of FE65. MCF 10A cells did not express APP or ApoEr2 but did express Dab1, X11α, and FE65. B) MCF 10A cells were cotransfected with ApoEr2 and X11α and treated with either control (lanes 1 and 3) or Reelin (lanes 2 and 4) for 24 h (lanes 1 and 2) or 20 min (lanes 3 and 4). Cell lysates were immunoprecipitated with anti-Flag for X11α and Western blotted for ApoEr2. Reelin decreased coprecipitation between ApoEr2 and X11α at 20 min (41%), but increased coprecipitation at 24 h (223%). C) MCF 10A cells were cotransfected with GFP-ApoEr2 and either vector (lanes 1, 3, and 5) or X11α (lanes 2, 4, and 6). Cell-surface proteins were biotin labeled, isolated with avidin beads, and immunoblotted with GFP for ApoEr2. Cell lysates showed similar levels of total ApoEr2. D) Quantification of data in C showed that full-length X11α increased surface levels of ApoEr2 by 64% (n=3) *P < 0.05.