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. 2021 Oct 22;6(20):e149651. doi: 10.1172/jci.insight.149651

Figure 4. TRAM deficiency gives rise to resolving monocytes with elevated expression of CD200R both in vitro and in vivo.

Figure 4

(A and B) Dimensionality reduction and clustering through uniform manifold approximation and projection (UMAP) of the scRNA-Seq data from WT (A) or TRAM monocytes (B) challenged with either PBS or 100 pg/mL LPS as we described in Methods. (C) Heatmaps showing representative genes differentially expressed in different clusters of monocytes challenged with subclinical-dose LPS. (D) Dot plot comparison of representative genes differentially expressed among LPS-treated WT versus Tram−/− monocytes. (E) BM cells from WT C57 BL/6 mice and Tram−/− mice were cultured with M-CSF (10 ng/mL) for 5 days. The surface expression of CD200R was analyzed and quantified by flow cytometry. (F) Apoe−/− Tram+/+ mice and Apoe−/− Tram−/− mice were fed with HFD for 8 weeks. Surface expression of CD200R on CD11b+Ly6C++ monocytes in the peripheral blood, BM, spleen, and aorta was examined by flow cytometry. Data are representative of 2 independent experiments, and error bars represent means ± SEM. *P < 0.05, and ***P < 0.001; Student’s 2-tailed t test (n = 3 to 10 for each group).