Strategy for determining proteomic consequences of NPM/ALK expression. Jurkat cells transfected with NPM/ALK cDNA were compared with those that were transfected with control LacZ (vector). One milligram from each cell lysate was labeled with either the light cICAT reagent (Lac Z Jurkat) or the heavy cICAT reagent (NPM/ALK-transfected Jurkat). The labeled proteins were combined; digested; subjected to cation-exchange chromatography, avidin chromatography, and reversed-phase chromatography; and analyzed by MS/MS. Selected proteins were verified by Western blot analysis, immunofluorescence microscopy, and immunohistochemistry and cellular pathways were functionally validated.