Figure 5.
Membrane-associated macrophages are controlled by CSF1 produced by stromal cells of serous membranes. (A) Whole-mount images of the two distinct membrane-associated macrophages and their quantification in Lyve1Cre:Csf1rfl/fl mice and littermate Csf1rfl/fl control mice. Scale bar, 50 µm. Data are representative of three independent experiments (n = 3 per genotype; mean ± SEM). Macrophages were quantified in multiple regions of mesenteric membrane per mouse. Unpaired Student’s t test: ****, P < 0.0001. (B) Expression pattern of Csf1, Il34, and Wt1 depicted on the t-SNE plot derived from scRNA-seq of whole mesenteric cells shown in Fig. 3 D. (C) Whole-mount images stained for LYVE1 in mesenteric membranes from WT1cre:R26LSL-tdTomato mice. Representative images of three independent experiments. Arrows indicate LYVE1lo/− macrophages (red, Wt1 Tomato reporter; green, LYVE1; blue, DAPI). Scale bar, 20 µm. (D) Whole-mount images of mesenteric membranes from Wt1Cre:Csf1fl/fl mice and littermate Csf1fl/fl control mice. Representative images of two independent experiments. Scale bar, 100 µm. (E) Quantification of LYVE1hi and LYVE1lo membrane-associated macrophages obtained from whole-mount images of Wt1Cre:Csf1fl/fl mice and littermate Csf1fl/fl control mice. Data are pooled from two independent experiments (Csf1fl/fl mice, n = 4; Wt1Cre:Csf1fl/fl mice, n = 6; mean ± SEM). Macrophages were quantified from multiple regions of mesenteric membrane per mouse. Unpaired Student’s t test: **, P < 0.01; ****, P < 0.0001. (F) Quantification of ICAM2+ macrophages in peritoneal cavity. Data are representative of at least three independent experiments (Csf1fl/fl mice, n = 4; Wt1Cre:Csf1fl/fl mice, n = 5; mean ± SEM). Unpaired Student’s t test: ****, P < 0.0001. (G) Quantification of Ly6Chi monocytes and neutrophils in blood. Data are representative of three independent experiments (Csf1fl/fl mice, n = 5; Wt1Cre:Csf1fl/fl mice, n = 4; mean ± SEM). Unpaired Student’s t test. (H) Confocal image from the mesenteric membrane of Adiponectincre: R26LSL-DTA mice and controls. Images are representative of two independent experiments. Scale bar, 50 µm.

Membrane-associated macrophages are controlled by CSF1 produced by stromal cells of serous membranes. (A) Whole-mount images of the two distinct membrane-associated macrophages and their quantification in Lyve1Cre:Csf1rfl/fl mice and littermate Csf1rfl/fl control mice. Scale bar, 50 µm. Data are representative of three independent experiments (n = 3 per genotype; mean ± SEM). Macrophages were quantified in multiple regions of mesenteric membrane per mouse. Unpaired Student’s t test: ****, P < 0.0001. (B) Expression pattern of Csf1, Il34, and Wt1 depicted on the t-SNE plot derived from scRNA-seq of whole mesenteric cells shown in Fig. 3 D. (C) Whole-mount images stained for LYVE1 in mesenteric membranes from WT1cre:R26LSL-tdTomato mice. Representative images of three independent experiments. Arrows indicate LYVE1lo/− macrophages (red, Wt1 Tomato reporter; green, LYVE1; blue, DAPI). Scale bar, 20 µm. (D) Whole-mount images of mesenteric membranes from Wt1Cre:Csf1fl/fl mice and littermate Csf1fl/fl control mice. Representative images of two independent experiments. Scale bar, 100 µm. (E) Quantification of LYVE1hi and LYVE1lo membrane-associated macrophages obtained from whole-mount images of Wt1Cre:Csf1fl/fl mice and littermate Csf1fl/fl control mice. Data are pooled from two independent experiments (Csf1fl/fl mice, n = 4; Wt1Cre:Csf1fl/fl mice, n = 6; mean ± SEM). Macrophages were quantified from multiple regions of mesenteric membrane per mouse. Unpaired Student’s t test: **, P < 0.01; ****, P < 0.0001. (F) Quantification of ICAM2+ macrophages in peritoneal cavity. Data are representative of at least three independent experiments (Csf1fl/fl mice, n = 4; Wt1Cre:Csf1fl/fl mice, n = 5; mean ± SEM). Unpaired Student’s t test: ****, P < 0.0001. (G) Quantification of Ly6Chi monocytes and neutrophils in blood. Data are representative of three independent experiments (Csf1fl/fl mice, n = 5; Wt1Cre:Csf1fl/fl mice, n = 4; mean ± SEM). Unpaired Student’s t test. (H) Confocal image from the mesenteric membrane of Adiponectincre: R26LSL-DTA mice and controls. Images are representative of two independent experiments. Scale bar, 50 µm.

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