Figure 2.
Anatomical niche impacts GPR183 expression by lung macrophages. (A)Gpr183-GFP expression by interstitial and alveolar macrophages from lung and BAL fluid of Gpr183GFP/+ (green histograms) and Gpr183+/+ mice (grey histograms), respectively. Gating strategy is shown in Fig. S1 A. Data are representative of three independent experiments with a total of n = 5–6 mice per genotype. (B)Gpr183-GFP expression by subsets of interstitial lung macrophages from Gpr183GFP/+ (green histograms) and Gpr183+/+ mice (grey histograms). Gating strategy is shown in Fig. S1 A. Data are representative of three independent experiments with a total of n = 6 mice per genotype. (C) Number of lung monocytes and macrophages in Gpr183+/+ and Gpr183−/− mice. Gating strategy is shown in Fig. S1 C. Data are represented as mean ± SEM. ns, not significant by unpaired Student’s t test. Lung data are pooled from three independent experiments with a total of n = 8 mice per genotype. BAL data are pooled from two independent experiments with a total of n = 5 mice per genotype. (D) Generation of Gpr183GFP/+ or Gpr183GFP/GFP (CD45.2+) → B6 (CD45.1+) bone marrow chimeras. Gpr183+/+ or Gpr183+/− (CD45.2+) → B6 (CD45.1+) chimeras were used as negative controls. (E)Gpr183-GFP expression by the indicated CD45.2+ monocyte–derived macrophage populations from the lung of Gpr183GFP/GFP (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (green histograms) 3 wk after bone marrow transfer. CD45.2+ macrophages from Gpr183+/− (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (grey histograms) were used as a control. Gating strategy is shown in Fig. S1 B. Data are representative of two independent bone marrow chimera experiments with a total of n = 4 mice per genotype. (F)Gpr183-GFP expression by CD45.2+ monocyte–derived alveolar macrophages from the BAL fluid of Gpr183GFP/+ (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (green histograms) 5 and 12 wk after bone marrow transfer. Macrophages from Gpr183+/+ (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (grey histograms) were used as a control. Gating strategy is shown in Fig. S1 A. Data are representative of two independent bone marrow chimera experiments with a total of n = 2–4 mice per genotype and time point. (G) Experimental setup to generate bone marrow–derived macrophages in vitro. Bone marrow cells from Gpr183GFP/+ or Gpr183+/+ mice were cultured with either M-CSF or GM-CSF and TGFβ1 to generate generic macrophages or alveolar macrophage-like cells, respectively. (H)Gpr183-GFP expression by in vitro–generated macrophages from Gpr183GFP/+ bone marrow cells (green histograms). Macrophages differentiated from Gpr183+/+ bone marrow were used as a control (grey histograms). Gpr183-GFP expression by Ly6Chi monocytes (bone marrow input) is shown on the left. Data are representative of two independent experiments with a total of n = 2 mice per genotype and two technical replicates per experiment. Panels D and G were adapted from Servier Medical Art.

Anatomical niche impacts GPR183 expression by lung macrophages. (A) Gpr183-GFP expression by interstitial and alveolar macrophages from lung and BAL fluid of Gpr183GFP/+ (green histograms) and Gpr183+/+ mice (grey histograms), respectively. Gating strategy is shown in Fig. S1 A. Data are representative of three independent experiments with a total of n = 5–6 mice per genotype. (B)Gpr183-GFP expression by subsets of interstitial lung macrophages from Gpr183GFP/+ (green histograms) and Gpr183+/+ mice (grey histograms). Gating strategy is shown in Fig. S1 A. Data are representative of three independent experiments with a total of n = 6 mice per genotype. (C) Number of lung monocytes and macrophages in Gpr183+/+ and Gpr183−/− mice. Gating strategy is shown in Fig. S1 C. Data are represented as mean ± SEM. ns, not significant by unpaired Student’s t test. Lung data are pooled from three independent experiments with a total of n = 8 mice per genotype. BAL data are pooled from two independent experiments with a total of n = 5 mice per genotype. (D) Generation of Gpr183GFP/+ or Gpr183GFP/GFP (CD45.2+) → B6 (CD45.1+) bone marrow chimeras. Gpr183+/+ or Gpr183+/− (CD45.2+) → B6 (CD45.1+) chimeras were used as negative controls. (E)Gpr183-GFP expression by the indicated CD45.2+ monocyte–derived macrophage populations from the lung of Gpr183GFP/GFP (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (green histograms) 3 wk after bone marrow transfer. CD45.2+ macrophages from Gpr183+/− (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (grey histograms) were used as a control. Gating strategy is shown in Fig. S1 B. Data are representative of two independent bone marrow chimera experiments with a total of n = 4 mice per genotype. (F)Gpr183-GFP expression by CD45.2+ monocyte–derived alveolar macrophages from the BAL fluid of Gpr183GFP/+ (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (green histograms) 5 and 12 wk after bone marrow transfer. Macrophages from Gpr183+/+ (CD45.2+) → B6 (CD45.1+) bone marrow chimeras (grey histograms) were used as a control. Gating strategy is shown in Fig. S1 A. Data are representative of two independent bone marrow chimera experiments with a total of n = 2–4 mice per genotype and time point. (G) Experimental setup to generate bone marrow–derived macrophages in vitro. Bone marrow cells from Gpr183GFP/+ or Gpr183+/+ mice were cultured with either M-CSF or GM-CSF and TGFβ1 to generate generic macrophages or alveolar macrophage-like cells, respectively. (H)Gpr183-GFP expression by in vitro–generated macrophages from Gpr183GFP/+ bone marrow cells (green histograms). Macrophages differentiated from Gpr183+/+ bone marrow were used as a control (grey histograms). Gpr183-GFP expression by Ly6Chi monocytes (bone marrow input) is shown on the left. Data are representative of two independent experiments with a total of n = 2 mice per genotype and two technical replicates per experiment. Panels D and G were adapted from Servier Medical Art.

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