Lipolysis assay

Adipocytes were insulin and serum starved for 4 h at 37°C with warm 0.1% BSA phenol red free basal medium. Lipolysis was induced with 1 uM isoproterenol (Sigma-Aldrich, St. Louis, MO, USA; #I6504) for 2 h at 37°C in 3% BSA KRBH (135 mM NaCl, 5 mM KCl, 1 mM MgSO4, 0.4 mM K2HPO4, 1 mM CaCl2, 5.5 mM Glucose, 20 mM HEPES; pH 7.4) and assessed by glycerol concentrations in supernatants (Glycerol Assay Kit, Sigma-Aldrich, St. Louis, MO, USA; #MAK117), calibrated by cell DNA content (CyQUANT, Thermo Fisher Scientific Inc., Waltham, MA, USA; #C7026). Glycerol concentrations in non-treated wells were considered as basal values.