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. 2010 Jan 12;9(6):1047–1062. doi: 10.1074/mcp.M900486-MCP200

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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Fig. 1. — Stimulation conditions and experimental design. A, LPA- and HB-EGF-induced tyrosine phosphorylation of the EGFR in SILAC-encoded A498 cells. Immunoblot analysis of total cell extracts with antibody recognizing Tyr(P)¹¹⁷³ in the EGFR revealed similar stimulation upon treatment with either 0.5 ng/ml HB-EGF or 10 μm LPA. EGFR levels were similar as verified by immunoblotting with anti-EGFR antibody. B, schematic illustration of the integrated proteomics work flow for quantitative phosphorylation analysis of total lysates and kinase-enriched fractions. pY, phosphotyrosine.