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. 2007 Jun 25;27(17):6183–6194. doi: 10.1128/MCB.00132-07

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Copyright © 2007, American Society for Microbiology

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FIG. 7. — Erbin does not affect phosphorylation of Smad2/Smad3. (A and B) Erbin does not affect ALK5(T202D)-mediated phosphorylation of Smad2/Smad3. Cell lysates were prepared from HEK293T cells cotransfected with Flag-Smad2, Flag-Smad3, ALK5(T202D), and Myc-Erbin as indicated. Phosphorylation of Smad2/Smad3 was detected by Western blotting with anti-P-Smad2 (αP-Smad2) and anti-P-Smad3 (αP-Smad3) antibodies. Expression levels of Myc-Erbin or Flag-Smad2 and Flag-Smad3 were detected by anti-Myc (αMyc) or anti-Flag (αFlag) antibodies. (C and D) Erbin does not affect TGFβ-induced phosphorylation of Smad2/Smad3. Cell lysates were prepared from HEK293T cells that were cotransfected with Flag-Smad2 and Flag-Smad3 with or without Myc-Erbin and treated with or without TGFβ (1 h). Flag-Smad proteins first were immunoprecipitated by anti-Flag antibody and then were subjected to Western blot analysis. WCL, whole-cell lysate; IB, immunoblot; IP, immunoprecipitate.