Studying the subcellular localization and DNA-binding activity of FoxO transcription factors, downstream effectors of PI3K/Akt.
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.
Essafi
Abdelkader
A
Cancer Research UK Laboratories, Department of Oncology, Imperial College London, London, UK.
Gomes
Ana R
AR
Pomeranz
Karen M
KM
Zwolinska
Aleksandra K
AK
Varshochi
Rana
R
McGovern
Ursula B
UB
Lam
Eric W-F
EW
D001483:Base Sequence; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D011232:Chemical Precipitation; D047369:Chromatin Immunoprecipitation; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D021381:Protein Transport; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction; D013010:Sonication
D051858:Forkhead Transcription Factors; D009687:Nuclear Proteins; D004247:DNA; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt