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It is a key part of the regulatory mechanism for appetite and energy stores, acting via central and peripheral actions

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Immunoblotting Cell pellets were washed twice with ice-cold PBS prior to lysis in RIPA buffer (10 mM Tris-Cl pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% SDS) supplemented with protease inhibitors Sigma-Aldrich, 11836170001) and phosphatase inhibitors (Roche, 04906837001)