Fig. 2.

ALK, pALK expression, and the effect of ceritinib treatment on RTK and signaling protein activity in ARMS and ERMS cell lines. a Endogenous ALK and phosphorylated ALK (pALK) expression in ARMS (Rh30, Rh41) and ERMS (RD and Rh18) cell lines as determined by Western Blot ((p)ALK = 220/140 kDa, Tubulin = 52 kDa). Aska cells were used as a positive control (these cells show a deletion in ALK, leading to constitutive activity). b Phosphorylation of 28 RTKs and 11 signaling proteins was assessed after 24 h ceritinib treatment (2.5 μM) for both ARMS and ERMS cell lines by PathScan analysis. Untreated (−) and treated (+) cells were compared. The boxes represent the location of ALK, IGF1R and Src on the array. c Heatmap representing the relative phosphorylation (as described in materials and methods) of the examined RTKs and signaling proteins for each cell line. Boxes represent the RTKs/signaling proteins that were further examined. d Effect of ceritinib treatment on ALK activity in ARMS and ERMS cell lines (relative ALK phophorylation) as determined by PathScan analysis