Expression of Bcr-Abl proteins and activation of Akt, Erk, and STAT5 in vivo and in vitro.

Western blot analyses of spleen cell lysates from 2 vector mice (lanes 1,2), 3 wt Bcr-Abl mice (lanes 3-5), 3 Bcr-Abl/R1057K mice (lanes 6-8), and 3 Bcr-Abl/ΔSH2 mice (lanes 9-11) (A) and of lysates of NIH3T3 cells transduced with titer-matched viruses carrying vector alone (lane 1), bcr-abl/R1057K (lane 2), bcr-abl/ΔSH2 (lane 3), or wt bcr-abl (lane 4) (B). The NIH3T3 cells were serum-starved for 12 hours 2 days after transduction. Equal amounts of total lysates of either spleen cells or NIH3T3 cells were separated on 6% to 15% polyacrylamide gradient gels, transferred to nitrocellulose filters, and probed with antibodies as indicated. The fresh filters were first probed with an anti-Abl antibody (Ab-3) or the phospho-specific antibodies against activated Akt, Erk, and STAT5 and then stripped and reprobed with an anti-Dynamin antibody or antibodies against corresponding Akt, Erk, and STAT5 proteins, respectively.