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Proteins were then precipitated by addition of 500 l methanol (pre-cooled at 80 C), 400 l LCMS-grade water, and 100 l chloroform, and the mixture was vortexed and centrifuged (10,000 g, 10 min, 4C) to afford a protein disc at the interface of CHCl3 and aqueous layers

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Ex-4 exerts regulatory control over the PI 3 K/Akt/Foxo1 pathway, impeding the nuclear entry of Foxo1 and consequently enhancing the expression of insulin gene transcription factors PDX-1 and MafA