FIG. 5.

SAM domain-mediated dimerization is insufficient for EN transformation. (A) Soft agar data from cells expressing MSCV control, WT EN, A93D-EN-GFP alone, or V112E-EN-V5 alone or NIH 3T3 cells expressing both A93D-EN-GFP and V112E-EN-V5. (B) NTRK3 IP of all cell lines followed by anti-phosphotyrosine Western blotting (top panel). Only WT EN protein is phosphorylated. Protein expression of all constructs was assayed by IB with anti-GFP and -V5 antibodies (middle panels). The association of A93D-EN-GFP with V112-EN-V5 was confirmed by performing anti-V5 IP followed by anti-GFP Western blots (bottom panel). Equal protein levels were determined with an anti-Grb2 antibody.