Figure 3: LPS-induced IL-18 levels are regulated by LXR activation.
From: The nuclear receptor LXR modulates interleukin-18 levels in macrophages through multiple mechanisms
(a) BMDM were treated with vehicle (DMSO), GW3965 (1 μ mol/L) or LXR antagonist (LXR ant, 1 μ mol/L) for 24 hours with or without LPS (100 ng/ml) for the last 6 hours. IL-18 mRNA levels were analyzed by RT-qPCR. Values indicate expression normalized to cyclophilin and are presented relative to the expression in LPS-treated cells, set as 1. Values represent the mean ± SEM (3–4 different BMDM preparations). t-test: ***p ≤ 0.001, ns p > 0.05. (b) IL-18 mRNA expression in BMDM from WT and LXRα β −/− mice were analyzed by RT-qPCR. Shown is a representative experiment performed in triplicate (mean ± SD) t-test: *p ≤ 0.05, ns p > 0.05. (c) BMDM were treated as in A and then activated with ATP for the last 2 hours. Pro-IL-18 expression was analyzed by immunoblotting and Hsp90 levels were assayed as loading control. Shown is a representative experiment of three. (d) BMDM were treated as in A. Intracellular expression of IL-18 is shown as analyzed by immunoblotting. Samples shown were analysed on non consecutive lanes of the same blot. (e) BMDM were treated as indicated. Secreted IL-18 levels in culture supernatants were quantified by ELISA. Values represent mean ± SEM (n = 3). t-test: *p ≤ 0.05.
