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Induction of FGFR2 and FGFR3 mRNA and protein in EGFR inhibitor treated NSCLC cells.

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posted on 29.11.2010 by Kathryn E. Ware, Marianne E. Marshall, Lydia R. Heasley, Lindsay Marek, Trista K. Hinz, Paula Hercule, Barbara A. Helfrich, Robert C. Doebele, Lynn E. Heasley

A. Quantitative RT-PCR assay for FGFR2 and FGFR3 mRNAs. Total RNA was purified from the indicated cell lines following a 3-day treatment with 1 µM gefitinib and submitted to quantitative RT-PCR analysis of FGFR2, FGFR3 and GAPDH. (Cell lines with EGFR autocrine signaling or gain-of-function EGFR mutations: open bars; cell lines lacking EGFR signaling: grey bars) The data are presented as fold expression over DMSO treated cells following normalization for GAPDH mRNA. B–C. Cell lysates from the indicated NSCLC cell lines treated 3 days with or without 1 µM gefitinib or 2 µg/mL Erbitux were immunoblotted for FGFR2, FGFR3, EGFR and the α-subunit of the NaK-ATPase as a loading control. Densitometry of FGFR2, FGFR3 and EGFR expression was normalized relative to NaK-ATPase expression and is indicated under each immunoblot.