Figure 3.

Prevention of cytokine-induced NF-κB activation in RINm5F cells. (a) RINm5F cells were pretreated with the indicated concentrations of kazinol C or isokazinol D for 3 h, and followed by IL-1β and IFN-γ treatment. Following 24 h of incubation, the concentration of nitrite and the expression of iNOS mRNA and protein were determined. Each value is the mean±s.e.m. of three independent experiments (n=9). **P<0.01 versus vehicle-treated control; ^##P<0.01 versus cytokine-treated cells. (b) After 30 min, NF-κB DNA binding was analyzed by electrophoretic mobility shift assay, and the nuclear translocation of the p65 and p50 subunits and the level of cytoplasmic IκBα were determined by western blotting. β-actin and proliferating cell nuclear antigen (PCNA) were used as loading controls for cytosolic and nuclear proteins, respectively.