The daily dose for rats was 1 mg/ kg body weight for 4 weeks

Effect of CL (CL-316243) alone or in combination with ADA, PTX or WT on PI(3,4,5)P3 production were studied using rat epididymal adipocytes. [32P]orthophosphate-labeled adipocytes that were treated or not treated with ADA (2 U/ml, 10 min) or PTX (1 μg/ml, 60 min) were incubated for 1 min with or without CL (1 μM). In addition, [32P]orthophosphate-labeled adipocytes treated with or without WT (3 μM, 1 min) were incubated for 1 min with or without CL or INS (0.7 μM). Furthermore, adipocytes treated or not treated with ADA or PTX were incubated for 10 min with or without CL. The radioactivity of PI(3,4,5)P3 (PIP3) that were separated by TLC and the accumulation of cAMP in adipocytes were determined.