Fig. 5
Functional properties of IM subpopulations at steady-state and dynamic regulation after airway exposure to microbial products. a Experimental outline for (b). FACS-sorted IM subpopulations were cultured ex vivo overnight with or without LPS, and supernatants were subjected to proteome profiling. b Heatmap depicting the relative abundance of the indicated molecules in the supernatants of non-stimulated (NS) or LPS-stimulated (LPS) lung IM subpopulations. Data represent the mean and are representative of one of 2 independent sorting experiments. c Representative contour plots showing steady-state IL-10 expression as assessed by detection of 450 nm fluorescence (blue fluorescent product of the cleaved CCF4 substrate) in CCF4-loaded cells isolated from IL-10-β-lactamase reporter ITIB or WT control mice. Numbers indicate % of IL-10+ cells within the cell populations, as quantified in (d). d Percentages of IL-10+ cells in the indicated populations. e Experimental outline for panels (f–i). f Kinetic analysis of numbers of each mononuclear phagocyte subset after i.t. instillation of Pam3CSK4, LPS, and CpG. g Representative contour plot of CD16.2 and CD206 expression within CD64+ IM from control (Day 0), and Pam3CSK4-, LPS- and CpG-injected mice 7 days after treatment. h Kinetic analysis of MHC-II expression within the indicated populations after Pam3CSK4, LPS or CpG treatment. i Numbers of IL-10+ cells within the indicated subpopulations 7 days after treatment. d, f, h, i Data show mean ± s.e.m., as well as individual mice in (d, i), and are pooled from 2 independent experiments (d, n = 9; f, h, n = 5–6/time point; i, n = 3–5). P-values were calculated using d, i non-parametric Mann–Whitney tests for pairwise comparisons or f, h two-way ANOVA with Tukey’s post hoc tests. *P < 0.05; **P < 10−2; ***P < 10−3; ****/°°°°P < 10−4; ns, not significant. In f the empty circles compare numbers of CD206+ IM 3 days after LPS vs. day 0. Source data are provided as a Source Data file. i.t., intratracheal
