Materials

The chemicals for Western blotting were purchased from Amersham (Bucks, UK). RNAzol^TM B was purchased from TEL-TEST Inc. (Friendwood, TX, USA). Primary antibodies against FoxO1, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), transcription factor IIB (TFIIB), tyrosine reside phosphorylation of insulin receptor substrate (p-IRS (Tyr632)), p-IRS (Ser307), phosphorylation-serine/threonine protein kinase (p-Akt), total Akt, peroxisome proliferator-activated receptor-alpha (PPARα), PPARβ, PPARγ, β-arrestin, phosphorylation-AMP-activated protein kinase (p-AMPK), AMPK, sterol regulatory element-binding protein 1c (SREBP-1c), and PAR2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Primary antibodies against p-FoxO1 (Thr24) and p-FoxO1 (Ser256) were manufactured by Cell Signaling Technology, Inc. (Danvers, MA, USA). Secondary antibodies, both anti-rabbit and anti-mouse immunoglobulin G (IgG)-horseradish peroxidase-conjugated antibodies, were purchased from Amersham. Horseradish peroxidase-conjugated donkey anti-sheep/goat IgG was obtained from Serotec (Oxford, UK). Polyvinylidene difluoride (PVDF) transfer membranes were obtained from manufacturer Millipore Corporation (Bedford, MA, USA).