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Cells were lysed for 20 minutes in buffer containing: 50 mM HEPES pH 7.4, 40 mM NaCl, 2 mM EDTA, 0.5% Triton X-100, 1.5 mM sodium orthovanadate, 50 mM NaF, 10 mM sodium pyrophosphate, 10 mM sodium -glycerophosphate, and protease inhibitor tablet (Roche, Nutley, N.J.)

Consistent with our microarray data, we found a significant increase in the expression of Fasn , which encodes fatty acid synthetase, and Ppara , which encodes PPAR, the master transcriptional regulator for -oxidation, in adipocytes lacking GPR146 (Fig