Research Article

Integrated Functional Analysis of the Nuclear Proteome of Classically and Alternatively Activated Macrophages

Figure 1

Polarization of macrophages (Mφ). Mouse immortalized bone marrow-derived Mφ were treated with IFN-γ/LPS or IL-4 for 18 h. (a) Representative flow cytometry images of the polarized subsets gated for the surface expression of CD11b and F4/80 (Mφ markers, a) and then screened for increase in the expression of CD64 (b) and TNF-α (d) as the markers of classical/proinflammatory Mφ and for CD206 (c) and arginase I (e) as the markers of alternative/anti-inflammatory Mφ are shown. (b–e) Bar graphs show the median fluorescence intensity for the expression of CD64 (b), CD206 (c), TNF-α (d), and arginase I (e) in control (untreated) and IFN-γ/LPS- or IL-4-polarized Mφ after 18 h of incubation (-6 biological replicates per group per observation). (f) Cell supernatants were used to measure the release of nitric oxide by the Griess assay ( biological replicates per group, triplicate observations per sample). Data in bar graphs are presented as mean . Data were analyzed by using InStat version 3 (GraphPad, La Jolla, CA) and analyzed by the Student test (for comparison of 2 groups) or one-way analysis of variance (ANOVA) with Tukey’s post hoc test (for comparison of multiple groups). The significance was accepted at value ≤ 0.05 (nontreated vs. treated).
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