Figure 3. S-nitrosylation inhibits the interaction of β-arrestin 2 and eNOS.
(A) HEK-eNOS cells transfected with FLAG-tagged β-arrestin 2 were exposed to L-NAME (3mM) or D-NAME (3mM) for 48 h or untreated, and L-NAME-treated and untreated cells were exposed to isoproterenol (ISO; 10 µM) for 10 min or to Cys-NO (50 µM) for 20 min. Lysates were immunoprecipitated with anti-eNOS or anti-FLAG antibody, and IPs and lysates were immunoblotted with anti-FLAG antibody (β-Arr2) or anti-eNOS antibody. (B) The amounts of β-arrestin 2 bound to eNOS and eNOS bound to β-arrestin 2 are represented as percentages of control values, normalized with respect to the immunoprecipitated levels of eNOS and β-arrestin 2 respectively. Data are means ± SE (n = 3); *P < 0. 01 re. control.