Figure 3. The lipolytic pathway is unaltered by DPI treatment.

Human differentiated adipocytes were treated with or without 10 µM DPI in the absence or presence of 5 µM forskolin. (A) 15 minutes in KRB with 1% BSA and 0.5 mM oleate. Cell lysates were collected and assayed for cAMP using an ELISA-based assay. (B, C) Cells were treated under the same conditions for 4 hours. Individual proteins were analyzed by western blotting with specific antibodies. Signals were visualized with an enhanced chemiluminescence substrate kit (Thermo Scientific, Rockford IL) as described in materials and methods. Results for cAMP are normalized to the basal condition and represented as fold increase. Average basal value was 3.9 ± 0.5 pmols per million cells, average forskolin-stimulated value was 68.9 ± 13.9 pmols per million cells. Values ± S.E.M. for three independent experiments. Immunoblots shown are representative of those obtained in three separate experiments.