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. 2008 Jun 9;205(6):1269–1276. doi: 10.1084/jem.20080124

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© 2008 Fong et al.

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Figure 3. — IKKβ inhibits M1 macrophage activation in response to LPS in vivo. (A) Peritoneal macrophages (PM) and (B) neutrophils (PMN) were isolated from IKKβ^ΔMye and IKKβ^f/f mice for in vitro killing assays with GBS (MOI of 5:1). (C) IKKβ^ΔMye and IKKβ^f/f mice were challenged intranasally with 10 ng LPS in PBS. After 4 h, lungs were harvested and digested for FACS analysis of CD11b (myeloid), Gr-1 (PMN), and MHC class II expression. A representative histogram of MHC class II expression on CD11b⁺ Gr-1⁻ alveolar macrophages is shown. (D) Tabulated data of mean fluorescence intensity for CD11b⁺ Gr-1⁻ MHC class II⁺–activated macrophages in the lung and PMN recruitment as percentage of BAL cells from LPS-challenged mice (data are represented as mean ± SEM of n = 6). (E) In parallel experiments, IL-12 and IL-10 levels were measured in lung homogenates from LPS-challenged mice by ELISA. Data are represented as mean ± SEM (n = 8; *, P = 0.0381). (F) Immunohistochemical analysis of NOS2 expression in LPS-challenged mice. IKKβ^ΔMye mice show increased expression of NOS2 in alveolar macrophages compared with IKKβ^f/f control mice (arrows). Representative panels are shown from n = 6 mice. Bar, 100 μm.