Fig. 2

In vivo tumors derived from the ALKIma1 cell line. A. Western blot analysis of NPM-ALK, and phosphorylated NPM-ALK (P-NPM-ALK) in CD4+ NA cells, 6 days post-transfection (NA cells-D6) and the ALKIma1 cell line. GAPDH was used as a loading control. B. Cytospin analysis of primary activated T cells (5 days post-activation) and ALKIma1 cells. C. Karyotype analysis of ALKIma1 cells. Red arrows show the t(2;5)(p23;q35) translocation, blue arrows indicate additional chromosomal events. D. Flow cytometry assessment of CD4, CD8 and CD3, CD30 expression levels in ALKIma1 cells. E. Der2 and Der5 sequences obtained by Sanger sequencing of ALKIma1 cells, and analysis of TCRγ clonality via multiplex PCR of ALKIma1 cells. F. Western blot analysis of NPM-ALK, STAT3, phosphorylated NPM-ALK (P-NPM-ALK) and phosphorylated STAT3 (P-STAT3) in ALKIma1 cells treated with crizotinib for 48 h versus untreated ALKIma1 cells. Activated T lymphocytes were used as a negative control. Vinculin was used as a loading control. Crizo: crizotinib. G. Cell survival analysis of ALKIma1 cells treated with increasing doses of crizotinib for 24, 48, 72 and 96 h. H. Temporal evolution of tumor volume in mice injected with ALKIma1 cells (subcutaneous injection). I. Histologic analysis of ALKIma1 tumor cells in skin nodules: H&E staining, anti-ALK, anti-CD3, anti-CD4 and anti-CD30