Studying the subcellular localization and DNA-binding activity of FoxO transcription factors, downstream effectors of PI3K/Akt

Abdelkader Essafi, Ana R Gomes, Karen M Pomeranz, Aleksandra K Zwolinska, Rana Varshochi, Ursula B McGovern, Eric W-F Lam

Research output: Contribution to journalArticle (Academic Journal)

15 Citations (Scopus)

Abstract

This chapter describes methods for studying downstream events of the PI3K/Akt signaling cascade, focusing on the FoxO transcription factors. These approaches also represent alternative means for gauging the phosphoinositide-3 kinase/Akt activity. We describe protocols for the fractionation of cytoplasmic and nuclear protein extracts and for studying transcription factor DNA-binding activity in vitro and in vivo.

Original languageEnglish
Pages (from-to)201-11
Number of pages11
JournalMethods in Molecular Biology
Volume462
DOIs
Publication statusPublished - 2009

Keywords

  • Base Sequence
  • Cell Line, Tumor
  • Cell Nucleus
  • Chemical Precipitation
  • Chromatin Immunoprecipitation
  • Cytosol
  • DNA
  • Forkhead Transcription Factors
  • Humans
  • Intracellular Space
  • Nuclear Proteins
  • Phosphatidylinositol 3-Kinases
  • Polymerase Chain Reaction
  • Protein Transport
  • Proto-Oncogene Proteins c-akt
  • Signal Transduction
  • Sonication
  • Journal Article

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