FIGURE 7.

Mutation of the β-actin binding domain in eNOS protein prevents eNOS-β-actin association and hyperoxia-induced increase in eNOS activity in COS-7 cells. COS-7 cells transfected with wild type and mutant eNOS plasmids were exposed to normoxia or hyperoxia (95% oxygen) for 24 h. Then, eNOS-β-actin association (A and B), eNOS activity (C), NO (D), and peroxynitrite (E) were determined as described under “Experimental Procedures.” A is a representative immunoblot from three experiments. B is a bar graph showing the changes in the ratio of eNOS to β-actin in the immunoprecipitates. Results are expressed as mean ± S.D.; n = 3 experiments. *, p < 0.05 versus normoxia in wild-type group; #, p < 0.05 versus control (without uric acid).