Fig. 9.

Effects of HDLs on IL-1β-induced NF-κB activation and IL-1β-mediated iNOS and COX-2 mRNA expression. Panel A, MIN6 cells were transfected with a NF-κB-driven firefly luciferase reporter and a Renilla luciferase-encoding construct. The following day, the cells were incubated 18 h with and without 10 ng/ml IL-1β in the absence or in the presence of 1 mm HDLs. NF-κB activity was then measured as described in Materials and Methods. The results correspond to the mean ± sem of five independent experiments. The threshold of significance was at P = 0.0167 (three comparisons). Panels B and C, MIN6 cells were treated or not with 10 ng/ml IL-1β for 6 or 24 h in the absence or in the presence of 1 mm HDLs. RNA was then extracted and reverse transcribed, and real-time PCR was performed to measure iNOS (panel B) and COX-2 (panel C) mRNA expression. The results are normalized to a control gene (RPLP0) and expressed as the percentage of the response obtained with a 6-h stimulation with IL-1β (mean ± sem of three and five independent experiments for the 6-h and 24-h time points, respectively). The threshold of significance was at 0.0125 (four comparisons). NS, Nonsignificant; C, control untreated cells.