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Testing of allosteric inhibition with probe 1a. (A) Dose–response curves with the allosteric inhibitor 4EGI-1. Assay conditions: 10 nM 1a, 100 nM eIF4E, and 50 mM HEPES buffer containing 100 mM KCl, 0.5 mM EDTA, and 1 mM DTT (pH 7.2). (B) Binding experiment in the absence/presence of 100 μM 4EGI-1. Assay conditions: 10 nM 1a and 50 mM HEPES buffer containing 100 mM KCl, 0.5 mM EDTA, and 1 mM DTT (pH 7.2).
