FIGURE 9.

Increased ATGL is recruited to LDs in WAT but not BAT following stimulation of tissues with a β-adrenergic receptor agonist. A, 100 mg of WAT and BAT from overnight-fasted mice were incubated for 2 h with adenosine deaminase (1 unit/ml) and phenylisopropyladenosine (100 nm) (basal conditions) or with adenosine deaminase (1 unit/ml) and isoproterenol (10 μm) (stimulated conditions) prior to measurement of glycerol content of media samples. B, fat cakes containing lipid droplets were isolated by ultracentrifugation from 100 mg of BAT and WAT lysates after incubations as described in A. Equal amounts of total fat cake protein were loaded as follows: for ATGL and CGI-58, 45 μg of total protein (same blot was re-used to identify ATGL and CGI-58); for plin5, 20 μg of total protein; and for plin 1, 5 μg of total protein were loaded. Experiments were performed twice.