Figure 4.
Sub-tissular location is a dominant determinant of small intestinal macrophage function. (A) Left: Representative flow cytometry plots showing the frequency of cells expressing pro-IL-1β from small intestinal cells enriched for macrophages and stimulated for 3 h. Right: Frequencies of stimulated macrophages expressing pro-IL1 β. (B) Left: Representative flow cytometry plots showing the frequency of cells expressing TNF-α from small intestinal cells enriched for macrophages and stimulated for 3 h. Right: Frequencies of stimulated macrophages expressing TNF-α. (C) Left: Representative flow cytometry plots showing the frequency of cells expressing iNOS from small intestinal cells enriched for macrophages and stimulated overnight. Right: Frequencies of stimulated macrophages expressing iNOS. (D) Left: Representative flow cytometry plots showing the frequency of pHrodo+ cells after 40-min incubation with pHrodo beads. Right: Geometric mean fluorescence intensity (gMFI) of pHrodo expression in macrophages incubated with beads. (A–D) Numbers in flow cytometry plots denote the percentages of cells within the gate. Error bars show mean ± SD. Data (n = 6 per group) are pooled from at least two independent experiments. Statistical comparisons were performed with a with one-way ANOVA, with Tukey’s multiple comparison test for parametric data and a Kruskal–Wallis test with Dunn’s multiple comparison test for nonparametric data. Significance is shown for comparisons between Tim-4+ and Tim-4− macrophages within the same compartments: *, P ≤ 0.05; **, P ≤ 0.01; and ****, P ≤ 0.0001; and between Tim-4+ and Tim-4− macrophages across compartments: #, P ≤ 0.01; ###, P ≤ 0.001. Refer to the image caption for details. Panel A: Four flow cytometry plots and a vertical bar graph showing pro-I L-1 beta expression in stimulated macrophage subsets across T i m-4 and C D 163 populations. Panel B: Four flow cytometry plots and a vertical bar graphs showing T N F alpha expression in stimulated macrophages across T i m-4 and C D 163 subsets. Panel C: Four flow cytometry plots and a vertical bar graph showing i N O S expression in macrophage subsets after stimulation across compartments. Panel D: Four flow cytometry plots and a vertical bar graph showing p H r o d o uptake and g M F I measurements indicating phagocytic activity in macrophage subsets.

Sub-tissular location is a dominant determinant of small intestinal macrophage function. (A) Left: Representative flow cytometry plots showing the frequency of cells expressing pro-IL-1β from small intestinal cells enriched for macrophages and stimulated for 3 h. Right: Frequencies of stimulated macrophages expressing pro-IL1 β. (B) Left: Representative flow cytometry plots showing the frequency of cells expressing TNF-α from small intestinal cells enriched for macrophages and stimulated for 3 h. Right: Frequencies of stimulated macrophages expressing TNF-α. (C) Left: Representative flow cytometry plots showing the frequency of cells expressing iNOS from small intestinal cells enriched for macrophages and stimulated overnight. Right: Frequencies of stimulated macrophages expressing iNOS. (D) Left: Representative flow cytometry plots showing the frequency of pHrodo+ cells after 40-min incubation with pHrodo beads. Right: Geometric mean fluorescence intensity (gMFI) of pHrodo expression in macrophages incubated with beads. (A–D) Numbers in flow cytometry plots denote the percentages of cells within the gate. Error bars show mean ± SD. Data (n = 6 per group) are pooled from at least two independent experiments. Statistical comparisons were performed with a with one-way ANOVA, with Tukey’s multiple comparison test for parametric data and a Kruskal–Wallis test with Dunn’s multiple comparison test for nonparametric data. Significance is shown for comparisons between Tim-4+ and Tim-4− macrophages within the same compartments: *, P ≤ 0.05; **, P ≤ 0.01; and ****, P ≤ 0.0001; and between Tim-4+ and Tim-4− macrophages across compartments: #, P ≤ 0.01; ###, P ≤ 0.001.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal