Journal: Nature

Article Title: Alternative transcription initiation leads to expression of a novel ALK isoform in cancer

doi: 10.1038/nature15258

Figure Lengend Snippet: a, Dose–response curves to crizotinib of Ba/F3 cells expressing the indicated ALK isoforms in the presence or absence of IL-3. Error bars, mean ± s.e.m.; n = 3 biological replicates. b, Representative immunoblots of ALKATI-expressing Ba/F3 cells treated with increasing concentrations of crizotinib for 2 h. See Supplementary Fig. 1 for uncropped blots. c, Normalized tumour volume in mice implanted with NIH-3T3 cells expressing the indicated ALK isoforms and treated with vehicle (n = 8 tumours) or crizotinib (n = 10 tumours). Error bars, mean ± s.e.m.; see also Source Data. d, H&E staining and immunohistochemistry of explanted ALKATI-expressing tumours 48 h after first crizotinib treatment. e, Normalized bioluminescence signal of ALKATI-expressing, luciferase-labelled NIH-3T3 tumours treated with vehicle or crizotinib. Error bars, mean ± s.e.m.; n = 8 tumours; see also Source Data associated with this figure. f, Quantitative mRNA ALK profiling of a metastatic melanoma (MM-382) compared to wild-type ALK, EML4–ALK, or ALKATI using Nanostring nCounter. g, H&E staining and ALK immunohistochemistry (inset) of MM-382. Scale bars in d and g, 50 μm. h, Computed tomography images of a subcutaneous melanoma metastasis from patient 1 (MM-382) before and after crizotinib treatment.

Article Snippet: For cell viability assays and ALK inhibitor–dose-response curves, 2,000 Ba/F3 cells were plated in triplicates in 96-well plates with increasing concentrations of the ALK inhibitors crizotinib (LC laboratories), TAE-684 (ChemieTek), or ceritinib (ChemieTek) as indicated.

Techniques: Expressing, Western Blot, Staining, Immunohistochemistry, Luciferase, Computed Tomography