NO-dependent effects of leptin on glyceroneogenesis and NOS III phosphorylation in WAT explants from rats.

Explants from SD (A) or Zucker rats (B, C) were pre-treated or not with L-NAME (1 mmol/L) or AG490 (10 µmol/L) for 30 min, then exposed or not to either leptin (10 µg/L), SNAP (1 mmol/L) or IFN-γ (50 µg/L) for 2 h in KRB medium containing 2% BSA and [1-¹⁴C]-pyruvate. Glyceroneogenic flux was measured by the [1-¹⁴C]-pyruvate incorporation into neutral lipids. Each value represents the mean ± SEM, (n = 4) *, P<0.01 vs. control; ^a P<0.01 vs. leptin-treated explants. (C) Representative autoradiogram of a western blot performed on WAT cytosolic proteins from Zucker rats reveals total NOS III and its Ser¹¹⁷⁹ phosphorylated form.