Design: Firstly, residues 244–476 of the FXR ligand binding domain (FXR-LBD) were cloned into Nde1/XhoI sites of pET-15b vector to get the plasmid expressing N-terminal His-FXR-LBD. His-FXR-LBD protein was purified according to the previously published method. Secondly, we used the AlphaScreen-Based Protein-Peptide Interaction assay to screen FXR antagonist from the compounds library. Furthermore, a transactivation assay was carried out to detect the antagonistic effect of the compound on FXR.
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