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

doi:10.1007/978-1-60327-115-8_13

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

Methods Mol Biol. 2009:462:201-11. doi: 10.1007/978-1-60327-115-8_13.

Authors

Abdelkader Essafi¹, Ana R Gomes, Karen M Pomeranz, Aleksandra K Zwolinska, Rana Varshochi, Ursula B McGovern, Eric W-F Lam

Affiliation

¹ Cancer Research UK Laboratories, Department of Oncology, Imperial College London, London, UK.

PMID: 19160671
DOI: 10.1007/978-1-60327-115-8_13

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.

MeSH terms

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

Substances

Forkhead Transcription Factors
Nuclear Proteins
DNA
Phosphatidylinositol 3-Kinases
Proto-Oncogene Proteins c-akt