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. 2004 Dec 6;167(5):945–952. doi: 10.1083/jcb.200404108

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Copyright © 2004, The Rockefeller University Press

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Figure 7. — Shank3 interacts with Grb2. (a) Schematic representation of the used Ret9 and Shank3 mutants. The consensus Grb2-binding motif in Shank3 is underlined. (b) Interaction of Shank3 with endogenous Grb2. HEK293 cells were cotransfected with Flag-tagged Shank3–Pro2 or Shank3–Pro2 containing the Y1006F mutant and Ret9, Ret9FA, or Ret9Y1062F. Immunoprecipitation of Shank3 using Flag–M2 Sepharose beads was performed followed by SDS-PAGE and Western blotting. Western blots of immunoprecipitations and total lysates were developed with antibodies against Grb2, Flag, phospho-tyrosine (PY), and Ret9 as indicated.