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Published in final edited form as: Nat Genet. 2012 Jul 1;44(8):852–860. doi: 10.1038/ng.2330

a, HCC827 ER1-ER5 sublines are resistant to erlotinib treatment as measured by CellTiterGLO cell viability assay. Data are from 3 independent experiments and are expressed as percent of vehicle treated cells and mean ± SEM. b, Expression of AXL and GAS6 in the ER sublines compared with parental HCC827 cells (data are from Western blot analysis). c, Erlotinib IC₅₀ in HCC827 cell lines (as indicated) measured 48h after treatment with a non-targeting or AXL or GAS6 siRNA. Erlotinib IC₅₀ is shown in parentheses. Data are representative of 3 independent experiments. d, Erlotinib IC₅₀ in HCC827 cell lines (as indicated) measured 48h after treatment with vehicle (control) or with MP-470 (1 µM) or XL-880 (1 µM) and erlotinib. Erlotinib IC₅₀ is shown in parentheses. Data are representative of 3 independent experiments. e, Effects of treatment for 48h with a non-targeting (−) or AXL siRNA in parental or ER1 and ER2 cell lines in the absence and presence of erlotinib on the indicated biomarkers. Data represent 3 independent experiments. f–g, Effects of treatment for 48h with a vehicle or the indicated doses of (f) MP-470 or (g) XL-880 in parental or ER1 and ER2 cell lines in the absence and presence of erlotinib on the indicated biomarkers. Data represent 3 independent experiments. h,Erlotinib IC₅₀ in HCC827 cells measured 5 days after transfection the cDNA constructs encoding the indicated proteins and treated with either vehicle (left) or with XL-880 (1 µM) and erlotinib. Erlotinib IC₅₀ is shown in parentheses. Data are representative of 3 independent experiments. i, Western blot for the indicated proteins in lysates from cells transfected with the indicated cDNA constructs and treated with XL-880 (1 µM) for 3 hours prior to cell lysis (h).

a, HCC827 ER1-ER5 sublines are resistant to erlotinib treatment as measured by CellTiterGLO cell viability assay. Data are from 3 independent experiments and are expressed as percent of vehicle treated cells and mean ± SEM. b, Expression of AXL and GAS6 in the ER sublines compared with parental HCC827 cells (data are from Western blot analysis). c, Erlotinib IC₅₀ in HCC827 cell lines (as indicated) measured 48h after treatment with a non-targeting or AXL or GAS6 siRNA. Erlotinib IC₅₀ is shown in parentheses. Data are representative of 3 independent experiments. d, Erlotinib IC₅₀ in HCC827 cell lines (as indicated) measured 48h after treatment with vehicle (control) or with MP-470 (1 µM) or XL-880 (1 µM) and erlotinib. Erlotinib IC₅₀ is shown in parentheses. Data are representative of 3 independent experiments. e, Effects of treatment for 48h with a non-targeting (−) or AXL siRNA in parental or ER1 and ER2 cell lines in the absence and presence of erlotinib on the indicated biomarkers. Data represent 3 independent experiments. f–g, Effects of treatment for 48h with a vehicle or the indicated doses of (f) MP-470 or (g) XL-880 in parental or ER1 and ER2 cell lines in the absence and presence of erlotinib on the indicated biomarkers. Data represent 3 independent experiments. h,Erlotinib IC₅₀ in HCC827 cells measured 5 days after transfection the cDNA constructs encoding the indicated proteins and treated with either vehicle (left) or with XL-880 (1 µM) and erlotinib. Erlotinib IC₅₀ is shown in parentheses. Data are representative of 3 independent experiments. i, Western blot for the indicated proteins in lysates from cells transfected with the indicated cDNA constructs and treated with XL-880 (1 µM) for 3 hours prior to cell lysis (h).