Fig. 1

Myeloid cells of LysM-Cre/Ikkβ ^F/F mice are relatively refractory to M1-like proinflammatory activation. a CD11b⁺ cells were isolated from the lumbar spinal cord of adult WT and LysM-Cre/Ikkβ ^F/F mice using CD11b microbeads. Genomic DNA from CD11b⁺ cells of each group was analyzed by real-time RT-PCR to determine the rate of ikkβ deletion. b Macrophages obtained from normal WT and LysM-Cre/Ikkβ ^F/F mice were stimulated for 6 h with lipopolysaccharide. Macrophages were isolated and analyzed to evaluate the degree of activation of TNF-α, IL-1β, IL-6, iNOS, IL-10, and TGF-β with real-time RT-PCR. The levels of gene expressions were compared to normal control. c-e Macrophages derived from WT and LysM-Cre/Ikkβ ^F/F mice were treated with lipopolysaccharide or IL-4, incubated with PE anti-mouse CD80 (c), APC anti-mouse CD86 (c), and APC anti-mouse CD206 (d), and used for flow cytometry analysis (c and d). Alteration in protein expression of M1 (iNOS) and M2 marker (Arginine-1) was analyzed by Western blot analysis (e). f-i Semi-thin sections from the lumbar spinal cord of adult WT (F and H) and LysM-Cre/Ikkβ ^F/F mice (g and i) were stained with toluidine blue. Panels h and i display high magnification micrographs of sections in (panels f and g) marked with squares, respectively. Bars = 100 μm. j-m Spinal cord lysate obtained from adult WT and LysM-Cre/Ikkβ ^F/F mice was analyzed for the expression of MBP by immunoblotting (j) and were quantified (k). The same preparation was analyzed for mRNA expression of MBP by real-time RT-PCR (l) and was quantified (m). Data are representative of 3 independent experiments with similar results. (ANOVA test; **p < 0.01 and *p < 0.05 versus WT mice; #p < 0.05 and ##p < 0.01 versus normal control mice)