Effect of compound 1h on NF-κB activation in LPS-activated macrophages.

<p>(<b>A</b>) RAW 264.7 macrophages or (<b>B</b>) J774A.1 macrophages (both 5 × 10⁵/ml; 1 ml) were incubated for 30 min with 10-40 µM compound <b>1h</b> or DMSO, then LPS (1 µg/ml) was added and incubation continued for 20 min, then levels of phosphorylated and total IKK-α and IκB-α were measured by Western blotting. (<b>C</b>) RAW 264.7 macrophages or (<b>D</b>) J774A.1 macrophages (both 5 × 10⁵/ml; 1 ml) were treated as in A and B, then nuclear translocation of NF-κB was analyzed by ELISA. (<b>E</b>) RAW-Blue^TM cells (5 × 10⁵/ml; 1 ml) were incubated for 30 min with 2.5-40 µM compound <b>1h</b> or DMSO, then LPS (1 µg/ml) was added and incubation continued for 24 h, then SEAP activity was measured by the QUANTI-Blue^TM assay and expressed as a percentage of that in the absence of compound <b>1h</b>. In (<b>A</b>) and (<b>B</b>), the results are representative of those obtained in three different experiments. In (<b>C</b>-<b>E</b>), the data are expressed as the mean ± SD for three separate experiments. *, **, and *** indicate a significant difference at the level of <i>p</i> < 0.05, <i>p</i> < 0.01, and <i>p</i> < 0.001, respectively, compared to the DMSO/LPS group.