Figure 3

Cell-type-specific effect of AZD0530. A BV173 (Ph+) and SEM (Ph-) cells were incubated with the indicated concentrations of AZD0530 and Imatinib for three days. Whole cell lysates were probed for activated SFKs, phosphorylated Hck and the activated form of Bcr-Abl. BV173 and WTSupB15 samples in the SEM (Ph-) panel were used as a control for the detection of Bcr-Abl and phosphorylated Bcr-Abl. Tubulin was used as control for protein loading. (1) In BV173 cells SFK activity but not Bcr-Abl activation was inhibited at concentrations that cause growth arrest and induce apoptosis. (2) In BV173 cells, SFKs were shown to act downstream of Bcr-Abl. B Bcr-Abl as substrate of SFKs. WTSupB15 (left) and RTSupB15 (right), were treated with AZD0530 and Imatinib for three days. Whole cell lysates were prepared and probed for activated SFKs, phosphorylated Hck and the activated form of Bcr-Abl. Tubulin was used as a control for equal protein loading. In both cell lines Bcr-Abl was shown to be transphosphorylated by SFKs.