Development of a novel recombinant cell line for detection and characterization of Ah receptor nuclear translocation in intact cells

Toxicol In Vitro. 2020 Aug;66:104873. doi: 10.1016/j.tiv.2020.104873. Epub 2020 Apr 23.


The Ah receptor (AhR) is a ligand-dependent transcriptional factor that mediates the effects of structurally diverse chemicals. Ligand binding stimulates nuclear translocation of the AhR and leads to AhR DNA binding and increased gene expression. Studies of the molecular mechanisms by which ligands bind to and activate the AhR and AhR-dependent signal transduction require methods to easily examine each step of the AhR signaling pathway. While current assays can measure ligand and DNA binding in vitro and gene expression in cells, there is no simple method to monitor AhR nuclear translocation. We developed a stably transfected mouse hepatoma cell line (yAHAYc6) that expresses yellow fluorescent protein-tagged AhR (yAhR) for use in qualitative or semiquantitative assessment of nuclear/cytoplasmic distribution of yAhR in living cells by fluorescent microscopy. yAhR nuclear translocation was stimulated in a concentration- and time-dependent manner by AhR agonists and inhibited by antagonists. Inhibition of nuclear export channels by leptomycin B, resulted in increased nuclear accumulation of yAhR in the absence of added ligand, indicating endogenous nucleocytoplasmic shuttling of unliganded AhR and demonstrating the utility of these cells. This novel cell line can be used to detect and characterize AhR ligands and will facilitate mechanistic studies of AhR signaling.

Keywords: AhR; Aryl hydrocarbon receptor; Nuclear translocation 2,3,7,8-tetrachlorodibenzo-p-dioxin; TCDD; YFP.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • COS Cells
  • Cell Line, Tumor*
  • Chlorocebus aethiops
  • Genes, Reporter
  • Ligands
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Plasmids
  • Polychlorinated Biphenyls / pharmacology
  • Polychlorinated Dibenzodioxins / pharmacology
  • Receptors, Aryl Hydrocarbon / agonists
  • Receptors, Aryl Hydrocarbon / antagonists & inhibitors
  • Receptors, Aryl Hydrocarbon / genetics
  • Receptors, Aryl Hydrocarbon / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Signal Transduction
  • Transfection


  • Bacterial Proteins
  • Ligands
  • Luminescent Proteins
  • Polychlorinated Dibenzodioxins
  • Receptors, Aryl Hydrocarbon
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria
  • Polychlorinated Biphenyls