The E-Morph Screening Assay: Phenotypic Screening for Estrogenic or Anti-estrogenic Compound Activities

The unintended exposure to compounds interfering with the estrogen hormone system can pose a threat to human health and the environment. In this chapter, we describe the E-Morph Screening Assay, a targeted imaging-based high-throughput phenotypic screening assay that allows the efficient identification and characterization of such compounds with estrogenic or anti-estrogenic activities in a 96- or 384-well plate format. It captures the phenotypic response of MCF-7 breast cancer cells, which comprise a complete, functional, and interconnected estrogen signaling pathway. The E-Morph Screening Assay measures the change of E-Cadherin-GFP signal intensity at adherens junctions as a quantitative readout for estrogen-dependent phenotypic changes in the morphology of cell-cell contacts.