This nuclear receptor assay system utilizes proprietary non-human cells engineered to provide constitutive, high-level expression of the full-length human estrogen receptor 1 (NR3A1), a ligand-dependent transcription factor commonly referred to as ER(alpha). INDIGO's reporter cells include the luciferase reporter gene functionally linked to an ER(alpha)-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in ER(alpha) activity. Luciferase gene expression occurs after ligand-bound ER(alpha) undergoes nuclear translocation, DNA binding, recruitment and assembly of the co-activators and accessory factors required to form a functional transcription complex, culminating in expression of the target gene. Unlike some other cell-based assay strategies, the readout from INDIGO's reporter cells demands the same orchestration of all intracellular molecular interactions and events that can be expected to occur in vivo. ER(alpha) reporter cells are prepared using INDIGO
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