FIG. 7.

Inhibition of IKK activity by 15dPGJ₂. Macrophages were incubated for 20 min with LPS (200 ng/ml) and IFN-γ (10 U/ml); after homogenization, the IKK complex was immunoprecipitated with anti-IKK2 Ab. Kinase activity was monitored in vitro for 10 min, using GST-IκBα(1-317) as the substrate and in the presence of the indicated concentrations of PGs and rosiglitazone. P(Ser³²)IκBα was detected using a specific anti-P(Ser³²)IκBα Ab. The membrane was reprobed with anti-IκBα Ab (A). The incorporation of [³²P]phosphate into GST-IκBα(1-54) or IκBα^S/A was determined as previously described (B). Results show the mean ± SEM of the band intensities, expressed with respect to the condition in the absence of PG and rosiglitazone. ∗, P < 0.01 with respect to the condition in the absence of addition.