LC-MS/MS profiling of NPM-ALK⁺ cells identifies a common set of Tyr-phosphorylated proteins. (A) Comparison of the datasets derived from the immunoaffinity (PhosphoScan) and conventional LC-MS/MS-based proteomic approaches (Table 1). (B) Immunoaffinity profiling of NPM-ALK⁺ ALCL cell lines compared with NPM-ALK-positive HEK-293T-Rex cells. (C) Comparative PhosphoScan performed on 6 different ALCL cell lines. (D) Tyrosine phosphorylated sites on the NPM-ALK protein are represented with a gray dot (9 of these were identified in all ALK⁺ cell lines); the star represents newly discovered sites. Tyrosine sites known to interact with IRS1, SHC, and PLC-g are indicated. (E) The phosphorylation status of selected proteins was assessed in ALCL cell lines after treatment with CEP14083 and detected by phospho-specific antibodies as indicated.