Expression and kinase activity of wt Bcr-Abl and the SH2 mutants.

(A) Equal amounts of total lysates of NIH3T3 cells transduced with titer-matched viruses carrying gfp alone (lane 1),bcr-abl (lane 2), bcr-abl/R1057K (lane 3), orbcr-abl/ΔSH2 (lane 4) were analyzed by immunoblotting with anti-Abl antibody, Ab-3. The positions of Bcr-Abl and the endogenous c-Abl are indicated. (B) Bcr-Abl (lane 1), kinase-deficient Bcr-Abl/K1176R (lane 2), Bcr-Abl/R1057K (lane 3), and Bcr-Abl/ΔSH2 (lane 4) were transiently expressed in 293T cells, immunoprecipitated with an anti-Abl antibody (Ab-3), and subjected to an in vitro kinase assay using substrate GST-Crk (i) or GST-10a (ii) as well as immunoblotting with the anti-Abl antibody (iii).