National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2022;2474:3-9. doi: 10.1007/978-1-0716-2213-1_1.
The estrogen-related receptor alpha (ERRα, NR3B1) is an orphan nuclear receptor which plays a role in endocrine disruption, energy homeostasis, and cancer prognosis. One of the unique features of this transcription factor is the interplay with its cofactors. For instance, certain modulators require the presence of proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) alongside ERRα. Therefore, identification of ERRα agonists and antagonists require examination of this nuclear receptor alone and together with PGC-1α. In this book chapter, we describe the step-by-step protocol of a multiplex luciferase assay designed to identify ERRα agonists, antagonists, and toxicity in one quantitative high-throughput screening assay using two different stable cell lines.
雌激素相关受体α（ERRα，NR3B1）是一种孤儿核受体，在内分泌干扰、能量稳态和癌症预后中发挥作用。这种转录因子的一个独特特征是与它的辅助因子相互作用。例如，某些调节剂需要与 ERRα 一起存在增殖物激活受体γ共激活因子 1α（PGC-1α）。因此，鉴定 ERRα 的激动剂和拮抗剂需要单独检查该核受体以及与 PGC-1α 一起检查。在本章中，我们描述了一种多重荧光素酶测定法的逐步方案，该方法旨在使用两种不同的稳定细胞系，在一种定量高通量筛选测定法中鉴定 ERRα 的激动剂、拮抗剂和毒性。
