Figure 1.
Two distinct macrophage populations coexist in the avascular regions of mesenteric membranes. (A) Whole-mount images of gut mesentery in adult WT mice. Square box indicates a region of avascular mesenteric membrane. (B and C) Whole-mount images of the avascular mesenteric membrane from tamoxifen-induced Csf1rCreERT2:R26LSL-tdTomato mice (B) and Lyz2Cre:R26LSL-tdTomato mice (C). Scale bar, 50 µm. (D) Whole-mount images of mesenteric membrane from tamoxifen-induced Csf1rCreERT2:R26LSL-tdTomato:CX3CR1gfp mice. CX3CR1-GFP cells (left), Csf1r-expressing Tomato+ cells (middle), and merged pictures (right) for two distinct macrophage populations. Scale bar, 50 µm. (E) Immunohistochemistry analysis of a whole-mount mesenteric membrane from CX3CR1gfp/+ mouse stained for LYVE1. CX3CR1gfp expression (left), LYVE1 (middle), and merged pictures (right) for two distinct macrophage subsets. Scale bar, 50 µm. (F) Quantification of LYVE1hi CX3CR1gfplo/− macrophages and LYVE1lo/− CX3CR1gfphi macrophages in mesenteric membranes. Data are representative of three independent experiments (n = 3; mean ± SEM). Macrophages were quantified in two different regions of mesenteric membrane per mouse. Unpaired Student’s t test: ****, P < 0.0001. (G) Immunohistochemistry analysis of a whole-mount mesenteric membrane from Lyz2Cre:R26LSL-tdTomato mice stained with LYVE1 and MHCII. Scale bar, 50 µm. (H) Flow cytometric analysis of membrane-associated macrophages isolated from gut mesentery with CD45, F4/80, CD64, LYVE1, and MHC II staining. SSC, side scatter. (I) Frequency of LYVE1hi membrane-associated macrophages and LYVE1lo/− membrane-associated macrophages from flow cytometric analysis (H). Data are pooled from two independent experiments (n = 9; mean ± SEM). Unpaired Student’s t test: ****, P < 0.0001. All imaging data are representative of at least three independent experiments.

Two distinct macrophage populations coexist in the avascular regions of mesenteric membranes. (A) Whole-mount images of gut mesentery in adult WT mice. Square box indicates a region of avascular mesenteric membrane. (B and C) Whole-mount images of the avascular mesenteric membrane from tamoxifen-induced Csf1rCreERT2:R26LSL-tdTomato mice (B) and Lyz2Cre:R26LSL-tdTomato mice (C). Scale bar, 50 µm. (D) Whole-mount images of mesenteric membrane from tamoxifen-induced Csf1rCreERT2:R26LSL-tdTomato:CX3CR1gfp mice. CX3CR1-GFP cells (left), Csf1r-expressing Tomato+ cells (middle), and merged pictures (right) for two distinct macrophage populations. Scale bar, 50 µm. (E) Immunohistochemistry analysis of a whole-mount mesenteric membrane from CX3CR1gfp/+ mouse stained for LYVE1. CX3CR1gfp expression (left), LYVE1 (middle), and merged pictures (right) for two distinct macrophage subsets. Scale bar, 50 µm. (F) Quantification of LYVE1hi CX3CR1gfplo/− macrophages and LYVE1lo/− CX3CR1gfphi macrophages in mesenteric membranes. Data are representative of three independent experiments (n = 3; mean ± SEM). Macrophages were quantified in two different regions of mesenteric membrane per mouse. Unpaired Student’s t test: ****, P < 0.0001. (G) Immunohistochemistry analysis of a whole-mount mesenteric membrane from Lyz2Cre:R26LSL-tdTomato mice stained with LYVE1 and MHCII. Scale bar, 50 µm. (H) Flow cytometric analysis of membrane-associated macrophages isolated from gut mesentery with CD45, F4/80, CD64, LYVE1, and MHC II staining. SSC, side scatter. (I) Frequency of LYVE1hi membrane-associated macrophages and LYVE1lo/− membrane-associated macrophages from flow cytometric analysis (H). Data are pooled from two independent experiments (n = 9; mean ± SEM). Unpaired Student’s t test: ****, P < 0.0001. All imaging data are representative of at least three independent experiments.

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