Fig A1.

Example of reverse transcriptase polymerase chain reaction (RT-PCR) detection of EML4-ALK in a primary tumor sample and cell lines (RT-PCR for GAPDH expression was also performed as a control for RNA extraction and cDNA synthesis). In this patient (PID5), (A) RT-PCR and (B) DNA sequencing demonstrate the presence of EML4-ALK variant 5 (Wong DW, et al: Cancer 115:1723-1733, 2009), in which exon18 of EML4 is fused to exon20 of ALK (lane 1). NCI-H2228 and NCI-H3122 have previously been shown to harbor EML4-ALK variants 3 and 1, respectively (Koivunen JP, et al: Clin Cancer Res 14:4275-4283, 2008). Variant 1, which results from a fusion of EML4 exon13 with exon20 of ALK, is also detected in this assay (lane 3), whereas variant 3, in which EML4 exon6 is fused to exon20 of ALK, is not (lane 2).