Fig. 5.

Effect of iNOS dimerization inhibitor, BBS-1, on iNOS half-life. HEK 293 cells stably expressing iNOS were incubated for 18 h in the presence or absence of 1 μM BBS-1. (A) Western blotting of cells lysates, using anti-iNOS antibody (Left). UD, presence of UD of iNOS. iNOS activity was evaluated by measuring nitrite accumulation in culture media (n = 3) (Right). (B) Pulse–chase analysis was performed in the presence or absence of BBS-1. iNOS was immunoprecipitated with anti-iNOS antibody. Eluted proteins were separated by SDS/PAGE, and bands, representing ³⁵S-labeled iNOS were quantitated to calculate iNOS half-life (Lower). Filled circles and open squares denote cells incubated in the absence or presence of BBS-1, respectively.