Complex formation of Gab2, Shc, and Crkl with NPM-ALK and p85 in Karpas299 cell lysates.
The 2 × 107 Karpas299 cells were subjected to immunoprecipitations with the anti-Gab1,-Gab2,-Shc,-Crkl, or –Cbl antibodies and subjected to anti-NPM-ALK or anti-p85 immunoblotting as described in the “Materials and methods.” Association of NPM-ALK with Gab1, Gab2, Shc, Crkl, and Cbl are demonstrated by the anti-NPM-ALK immunoblotting (A, left panel). Anti-p85 immunoblotting showed the complex formation of Gab2, Shc, and Crkl with p85 (B, left panel). A rabbit antimouse antibody (RαM) served as a negative control for the immunoprecipitation. Immunoblotting with anti-Gab1,-Gab2,-Shc,-Crkl, or -Cbl antibodies demonstrated the immunoprecipitated proteins (C).
