FIGURE 4.
Role of adaptor molecules and IL-1R1 in P. aeruginosa activated macrophages. Macrophages were derived from bone marrow cells isolated from C57BL/6, TIRAP−/−MyD88−/−, TRIF−/−, and IL-1R1−/− mice and stimulated with LPS or with antibiotic killed 19660 or 19660ΔfliC. A, Representative wells showing translocation of NF-κB to the nucleus. Macrophages were incubated with killed 19660ΔfliC, and at indicated time points, cells were fixed and immunostained with Ab to the p65 subunit of NF-κB. Data are indicative of two repeat experiments with triplicate wells. B, CXCL1/KC and CCL5/RANTES production by bone marrow macrophages stimulated 3 h with 19660ΔfliC. C, Bone marrow macrophages were incubated with LPS, 19660, or 19660ΔfliC. At indicated times, cells were harvested and processed for Western blot analysis using Abs to P-IκB, P-IRF3, and β-actin.