FIGURE 4.

Intestinal macrophages do not phosphorylate NF-κB p65 or translocate NF-κB p50 into the nucleus. A, intestinal macrophages and autologous blood monocytes were stimulated with smooth LPS (1 μg/ml) for 5, 15, 30, or 60 min, and NF-κB p65 phosphorylation was assessed by flow cytometry. B, nuclear transport of NF-κB p50 was determined by comparison of NF-κB p50 levels in nuclear cell extracts by ELISA after exposure to LPS (1 μg/ml). Levels of nuclear NF-κB p50 were significantly lower in intestinal macrophages than blood monocytes at each time point, p < 0.006. C, intestinal macrophages and autologous blood monocytes (10 × 10⁶/ml) were incubated for 0 or 30 min with smooth LPS (1 μg/ml), and extracts were analyzed for MyD88-dependent and -independent NF-κB from signal proteins by Western blot (A). D, macrophages were also analyzed for Smad signal proteins by Western blot. Data are from a representative experiment (n = 3) (**, p < 0.01; *, p < 0.05).