Fig. 1.
Intracellular NO measurement by HPLC analysis of DAF-2 T (triazolofluorescein). HAEC were pretreated with wortmannin (500 nM), Akt inhibitor (20 µM), or L-NIO (0.1 mM) before addition of DAF-2 (5 µM) and tandem treatments with GTN (10 nM) or VEGF (20 ng/ml). Cellular concentrations of the product DAF-2 T were analyzed by HPLC and calculated using a standard curve. Reported data are DAF-2 T concentration normalized to protein content in each sample. *P<0.05, **P<0.01.