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. Author manuscript; available in PMC: 2011 Jul 1.
Published in final edited form as: Chem Biol Drug Des. 2010 May 4;76(1):1–9. doi: 10.1111/j.1747-0285.2010.00986.x

Figure 5.

Figure 5

Testing for inhibition of EGF receptor dimerization by two methods. A) Chemical cross-linking assay. Cells were pre-incubated with 1% DMSO (lane 1 and 2) or 100 μM of different inhibitors (lane 3-6) prior to stimulation with 25 nM EGF (lane 2-6) for 5 minutes. All cells were then treated with 3 mM BS3. NSC11241 (lane 3) and NSC56452 (lane 5) significantly inhibit dimer formation, while Lane 4 and 6 show compounds that did not inhibit dimer formation. B) Luciferase fragment complementation. Cells stably expressing ΔC-EGFR-NLuc and ΔC-EGFR-CLuc were pre-treated with DMSO, the indicated concentrations of NSC56452 or 1 μg/ml cetuximab for 20 min in the presence of 0.6 mg/ml D-luciferin prior to the addition of 3 nM EGF. All assays were performed in quadruplicate. Data represent the change in photon flux between cells treated with or without EGF.