Fig. 1.

Chemical manipulation of the lipolytic pathway inhibits aP2 secretion. A: Canonical lipolytic pathway and inhibitors used to block steps in this pathway. B–D: Differentiated 3T3-L1 adipocytes were pretreated with the PKA inhibitor H89 (B), ATGL inhibitor Atglistatin (C), HSL inhibitor 76-0079 (D), or HSL-MAGL dual inhibitor CAY10499 (E), followed by IBMX stimulation. Secreted aP2 was measured by Western blot analysis of CM. Glycerol release into CM was measured to assess lipolysis and normalized to total protein content (shown below or at right in each panel). Atgli, Atglistatin; CAY, CAY10499; CL, cell lysate; CM, conditioned media; Cont, control; I, IBMX; 76, 76-0079. Statistical analysis was done using Student’s t-test. * P < 0.05, *** P < 0.001. Bars indicate SEM. Western blots are representative of at least three independent experiments.