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cas2408-sup-0001-FigureS1.epsimage/eps1494KFig. S1. Left: Mutant and wild-type epidermal growth factor receptor (EGFR) plasmids were mixed at various ratios, then the mixed plasmids were amplified as a PCR template for fragment analyses. Calibration curve between mixture ratio of the plasmids and proportion ratio of amplified fragments. Right: Electropherograms of the fragment analysis of the mixtures of mutant and wild-type EGFR plasmids.
cas2408-sup-0002-FigureS2.epsimage/eps2094KFig. S2. Data for A549 cells are shown. (A) Left: Electropherograms of the fragment analyses. Right: Relative proportions of wild-type epidermal growth factor receptor (EGFR) expression. (B) Left: Gefitinib sensitivity curves. Right: IC50 values of cells to gefitinib. (C) Western blot analysis for total and phosphorylated EGFR, AKT, and ERK.
cas2408-sup-0003-FigureS3.epsimage/eps1500KFig. S3. (A) Direct sequencing of amplified fragment of exon 19 in epidermal growth factor receptor (EGFR) gene from PC9 cell-derived genomic DNA. (B) Copy numbers of EGFR gene were determined by quantitative PCR.
cas2408-sup-0004-FigureS4.epsimage/eps1578KFig. S4. (A) mRNA expression level of epidermal growth factor receptor (EGFR) ligands including EGF, epiregulin, amphiregulin, and TGFα in PC9 cells by quantitative PCR. (B) Hypoxia-inducible factor-1α (HIF1α) or transforming growth factor-α (TGFα) expression in HCC2935, A549, and HCC827 cells by Western blotting. (C) TGFα or HIF1α knockdown by siRNA reversed hypoxia-induced resistance to gefitinib in HCC2935 cells.
cas2408-sup-0005-FigureS5.epsimage/eps1685KFig. S5. Proposed model depicting the involvement of wild-type epidermal growth factor receptor (EGFR), hypoxia-inducible factor-1α (HIF1α), and transforming growth factor-α (TGFα) in hypoxia-induced resistance to gefitinib in PC9 cells.

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