Figure 6.

Stimulation of lipolysis in adipocytes from Rimonabant-treated rats. Male Wistar rats (280–320 g) were treated with one dose of Rimonabant (30 mg/kg, p.o.; RIM) or vehicle. After 5 h under free access to food, epididymal adipocytes were prepared by collagenase digestion and washed and then incubated (3 h, 37 °C) in the absence or presence of CP55.940 (10 µM final conc.). Upper panel: The amounts of glycerol and FA released into the incubation medium were determined enzymatically. Means of determinations in triplicate. Middle and lower panels: The amounts of HSL and perilipin-1 associated with LD, which were prepared from the homogenates of the adipocytes by centrifugation through sucrose cushions, were determined. For this, proteins contained in the top fraction were extracted from lipids, then separated by SDS-PAGE and analyzed for HSL and perilipin-1 by immunoblotting. The images of a typical experiment with electrophoretic runs in quadruplicate with their quantitative evaluations are shown. Mean ± SD of two different adipocyte/LD preparations with incubations/measurement in quadruplicate. * p ≤ 0.05; no significant differences between absence and presence of CP55.940.