L ne bitartrate salt monohydrate

Adipocytes were stimulated with or without adenosine 5′-monophosphate sodium salt (AMP) (300 nM) (Sigma-Aldrich, A1752), inosine (300 nM) (Sigma-Aldrich, I4125), hypoxanthine (300 nM) (Sigma-Aldrich, H9377) or l-(−)-NE (+)-bitartrate salt monohydrate (1 µM) (Sigma-Aldrich, A9512) for 15 min. Afterwards, cells were quickly washed with PBS and lysed with 0.1 M HCl. Subsequently, samples were analysed using Direct cAMP ELISA Kit (Enzo, ADI-901-066), following the manufacturer’s instructions. Measurement of optical density was performed at 405 nm using a plate reader (Perkin Elmer).

(−)-EPI (+)-bitartrate salt, L-(-)-NE (+)-bitartrate salt monohydrate, and ICI 118,551 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ketamine (Imalgene 1000, Merial, Lisboa, Portugal) and xilazine (Rompum 2%, Bayer, Lisboa, Portugal).