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

Methods in molecular biology (Clifton, N.J.) (2009-01-24)
Abdelkader Essafi, Ana R Gomes, Karen M Pomeranz, Aleksandra K Zwolinska, Rana Varshochi, Ursula B McGovern, Eric W-F Lam
RÉSUMÉ

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.

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LY 294002, InSolution, ≥98%, 10 mM, reversible and specific inhibitor of PI 3-kinase