Biochemical characterization and localization of EGFRΔ768.

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Biochemical characterization and localization of EGFRΔ768. Biochemical characterization and localization of EGFRΔ768. A, biochemical characterization of EGFRΔ768. 25 μmol/L of erlotinib and 50 μg/mL of cetuximab were used for the inhibition experiment. No primary antibody was added for immunoprecipitation (IP) in the cetuximab-treated cell lysates. EGFR phosphorylation/expression was measured as described previously (11). The quantification was depicted as a ratio below the gel. Control IP with IgG antibody were negative (data not shown). B, localization of EGFRΔ768-GFP in the NIH3T3 clonal cells. Solid white arrows pointed to GFP concentrated in the membrane ruffles, whereas hollow white arrows pointed to GFP concentrated at the cell to cell contact. C, cell surface expression of EGFRΔ768-GFP in 3T3 cells by cell fractionation and organelle isolation experiment. Noted the strong GFP signal in the plasma membrane fraction (left). The identity and relative purity of each fraction was confirmed by the expression of organelle specific markers (right). James Keller et al. Mol Cancer Res 2016;14:740-752 ©2016 by American Association for Cancer Research