Nilotinib effect on ABL and SRC tyrosine kinase activity.

<p><b>(A)</b> Triton X-100 lysates of BT-549 and MDA-MB 231 cells incubated with DMSO (-) or 100nM nilotinib (+) were used for <i>in vitro</i> kinase assays to test ABL kinase activity. Immunoprecipitates were incubated with GST-Crk as a substrate and P³²-labeled ATP. P³² incorporation by GST-Crk was detected by autoradiography. The Abl content of immunoprecipitates and densitometry quantification normalized to Abl are also shown. <b>(B)</b> Triton X-100 lysates of BT-549 and MDA-MB 231 cells incubated with DMSO (-) or 100nM nilotinib (+) were immunoprecipitated with an anti-CST1 antibody that recognizes SRC, FYN and YES. <i>In vitro</i> kinase assay to test SRC activity was performed using enolase as substrate and P³²-labeled ATP. P³² incorporation by enolase was detected by autoradiography. Src and tubulin are used as loading control. Densitometry quantification normalized to tubulin is shown.