High-Throughput Image-Based Screening to Identify Chemical Compounds Capable of Activating FOXO

Methods Mol Biol. 2019:1890:151-161. doi: 10.1007/978-1-4939-8900-3_13.

Abstract

FOXO proteins are transcription factors with important roles in the regulation of the expression of genes involved in cell growth, proliferation, differentiation, and longevity. FOXO proteins are active in the nucleus but, upon post-translational modification they form a docking site for 14-3-3 proteins and are translocated to the cytoplasm where they are inactive.We make use of this regulatory mechanism of FOXO proteins to develop an image-based high-throughput screening platform to detect compounds that regulate FOXO3 subcellular localization. This system has proven a powerful tool to isolate inhibitors of proteins upstream of FOXO, such as PI3K inhibitors.

Keywords: Forkhead box O proteins–FOXO; High-content screening; Nucleus-cytoplasm translocation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Data Analysis
  • Drug Discovery / methods*
  • Fluorescent Antibody Technique
  • Forkhead Transcription Factors / genetics*
  • Forkhead Transcription Factors / metabolism
  • Genes, Reporter
  • High-Throughput Screening Assays*
  • Humans
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Small Molecule Libraries
  • Transcriptional Activation / drug effects*

Substances

  • Forkhead Transcription Factors
  • Recombinant Fusion Proteins
  • Small Molecule Libraries