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After incubation, the conditioned medium was collected and the concentration of GLP and PYY was determined by enzyme immunoassay with a specific ELISA kits for GLP-1 and PYY (Peninsula Laboratories, LLC, San Carlos, CA) (Katsuma et al., Assay of phosphoinositide (PI) hydrolysis PI hydrolysis was measured as total inositol phosphate formation with the use of anion exchange chromatography as described previously (Murthy and Makhlouf, 3 H]inositol (0.7 Ci/ml) in inositol-free medium and treated with OA or 8-pCPT-2- O -Me-cAMP (10 M) in the presence or absence of inhibitors for 1 min in a medium containing 25 mM HEPES, 115 mM NaCl, 5.8 mM KCl, 2.1 mM KH 2 PO 4 , 2 mM CaCl 2 , 0.6 mM MgCl 2 , and 14 mM glucose

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Quantitative real-time PCR RNA was purified by using ReliaPrep RNA (Promega, WI, USA) and reverse transcription was performed using 1.5 g total RNA as template with the ImProm-II Reverse Transcription System (Promega, WI, USA)