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

Figure 4.

Figure 4.

Reelin decreases Fyn- and X11α-mediated tyrosine phosphorylation of ApoEr2 and decreases Fyn-mediated interaction between ApoEr2 and X11α. A) COS7 cells were transfected with ApoEr2 and vector, X11α, Fyn, or both X11α and Fyn for 24 h, and treated with control or Reelin-conditioned medium for 20 min. Cell lysates were immunoprecipitated for phosphotyrosine and Western blotted for ApoEr2 (α-HA) to detect tyrosine phosphorylated ApoEr2. B) Quantification of data shows that ApoEr2, Fyn, and X11α increased phospho-ApoEr2 levels by 186% (lane 4) compared to ApoEr2 and X11α (P<0.01) (lane 2) and by 109% compared to ApoEr2 and Fyn (P<0.05) (lane 3). Reelin treatment of ApoEr2 and Fyn transfected cells increased phospho-ApoEr2 by 214% (P<0.01) (lane 7), and Reelin treatment of cells triply transfected with X11α, ApoEr2, and Fyn decreased phospho-ApoEr2 by 47% (P<0.05) (lane 8). C) COS7 cells were transfected with ApoEr2, X11α, and vector (lane 1) or Fyn (lane 2), and treated with control medium, or transfected with ApoEr2, X11α, and Fyn and treated with Reelin for 20 min (lane 3). Lysates were immunoprecipitated with anti-Flag for X11α and Western blotted with anti-HA for ApoEr2 (top panel). Fyn increased the interaction between ApoEr2 and X11α by 153% (lane 2, P<0.05), and Reelin treatment reversed this effect (by 42%; lane 3 vs. lane 2). Bottom panels show consistent expression patterns.