“…[61] Because of their structural similarity to UDCA and other GR ligands, we tested the potential affinity of compounds (5, 6, 6a, 6b, 10, 11, 14, 16, 18, 19, 19a) for GR using a yeast-based fluorescence assay optimized for screening steroidal derivatives with estrogen, androgen or glucocorticoid receptor binding properties. [23,24] For GR expression in yeast, a galactose-inducible plasmid (pRF4-6-rGR-LBD-EYFP) encoding the ligand binding domain (LBD) of the glucocorticoid receptor (GR) fused to enhanced yellow fluorescent protein (EYFP) was used. Expression of GR-LBD-EYFP in yeast was detected by fluorescence microscopy, and dose-dependent binding of a positive control GR ligand, prednisolone, was confirmed.…”