Fig. 5
EML4-ALK promotes anchorage-independent growth in hTERT-immortalized normal human bronchial epithelial cells. a HBET1, an hTERT-immortalized normal human bronchial epithelial cell line, was transduced with the EML4-ALK lentiviral vector or the control vector as in Fig. 3. The cells at 28 days after transduction were examined in western blot analysis as in Figs. 3a and 4. Full-length blots/gels are presented in Supplementary Figure. b Cell proliferation curves of HBET1 expressing EML4-ALK or with the control vector. Cumulative PDL were calculated and plotted to days after transduction, as above. c Anchorage-independent growth of HBET1 cells expressing EML4-ALK. The HBET1 cells with EML4-ALK or the control vector were examined for anchorage-independent growth, as in Fig. 3d and S5C. Data analysis and presentation are also as in Fig. 3d and S5C. Arrows in a representative image indicate colonies that formed in soft-agar medium. Scale bars, 50 μm. ** P < 0.01. d Summary of tumorigenicity assay in NOD.SCID/Ncr mice. Cells were subcutaneously injected into each flank (5 × 106 per flank) of mice at 6–10 weeks of age, followed by observation until 8 weeks after injection. In each mouse, one flank had EML4-ALK-expressing cells or soft-agar clones, and the other flank had cells with the control vector. Whereas NIH/3 T3 cells expressing EML4-ALK (2 × 106 per flank, at both flanks of two mice) produced tumors necessitating euthanasia, no progressively growing tumors formed from any of hTERT-CRL-2097- and HBET1-derived cells