High content imaging-based assay to classify estrogen receptor-α ligands based on defined mechanistic outcomes

Gene. 2011 May 15;477(1-2):42-52. doi: 10.1016/j.gene.2011.01.009. Epub 2011 Jan 20.


Estrogen receptor-α (ER) is an important target both for therapeutic compounds and endocrine disrupting chemicals (EDCs); however, the mechanisms involved in chemical modulation of regulating ER transcriptional activity are inadequately understood. Here, we report the development of a high content analysis-based assay to describe ER activity that uniquely exploits a microscopically visible multi-copy integration of an ER-regulated promoter. Through automated single-cell analyses, we simultaneously quantified promoter occupancy, recruitment of transcriptional cofactors and large-scale chromatin changes in response to a panel of ER ligands and EDCs. Image-derived multi-parametric data was used to classify a panel of ligand responses at high resolution. We propose this system as a novel technology providing new mechanistic insights into EDC activities in a manner useful for both basic mechanistic studies and drug testing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Benzhydryl Compounds
  • Blotting, Western
  • Cell Line, Tumor
  • Cluster Analysis
  • Endocrine Disruptors / pharmacology
  • Estradiol / analogs & derivatives
  • Estradiol / pharmacology
  • Estrogen Antagonists / pharmacology
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / metabolism*
  • Fulvestrant
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence / methods*
  • Phenols / pharmacology
  • Principal Component Analysis
  • Promoter Regions, Genetic / genetics
  • Raloxifene Hydrochloride / pharmacology
  • Reproducibility of Results
  • Tamoxifen / analogs & derivatives
  • Tamoxifen / pharmacology


  • Benzhydryl Compounds
  • Endocrine Disruptors
  • Estrogen Antagonists
  • Estrogen Receptor alpha
  • Phenols
  • Tamoxifen
  • Green Fluorescent Proteins
  • afimoxifene
  • Fulvestrant
  • Raloxifene Hydrochloride
  • Estradiol
  • bisphenol A