FIGURE 4.

MGL and HSL are involved in MG degradation in WAT. a and b, basal and forskolin-stimulated release of FFA and glycerol from gonadal WAT. Fat pads (∼20 mg) were preincubated in Dulbecco's modified Eagle's medium, containing 2% defatted BSA in the absence or presence of 10 μm forskolin for 1 h at 37 °C. Thereafter, WAT pieces were transferred into identical fresh medium, and FFA and glycerol release were determined after incubation for another hour (n = 5 for each genotype). c, MGH activity in WAT was determined in 20,000 × g infranatants using various concentrations of rac-1(3)-OG as substrate. Experiments were performed in the absence or in the presence of the HSL inhibitor 76-0079 (n = 5 for each genotype). d, MGH activities were inhibited in 20,000 × g infranatants of WAT of wild-type mice using various concentrations of specific inhibitors for MGL (JZL 184), HSL (76-0079), or both inhibitors in combination (JZL184/76-0079). e, activity of HSL was determined in post-nuclear lysates (1000 × g supernatant) of COS-7 cells expressing murine HSL using 2-AG, rac-1(3)-OG, or 2-OG as substrate. The MGH activity detected in cells expressing β-galactosidase was set as blank. Data are presented as mean ± S.D. and are representative for at least three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001.