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A – C Integrating scRNA-seq analyses of Dlx5 + lineage cells, <t>Fgfr3</t> + lineage cells, Col1a1(2.3 kb)-GFP-marked osteoblasts, Cxcl12-GFP-marked CAR cells at P49 (pulsed at P21). A Diagram for LIGER data integration. scRNA-seq datasets of sorted GFP + and/or tdTomato + single cells harvested from Col1a1(2.3 kb)-GFP ; Dlx5-creER ; R26R tdTomato femur bone marrow at P21 (2920 cells) and Cxcl12-GFP <t>;</t> <t>Fgfr3-creER</t> ; R26R tdTomato (4347 cells) were merged by LIGER. Scale bar: 500 μm (whole images) or 20 μm (magnified view). B Left panel: UMAP plot of major classes of GFP + and/or tdTomato + cells (Cluster 0-12, 7267 cells). Center panels: Density plots of Col1a1(2.3 kb)-GFP ; Dlx5-creER ; R26R tdTomato (upper) and Cxcl12-GFP ; Fgfr3-creER ; R26R tdTomato derived cells (lower). Right panels: Feature plots of representative genes enriched in each cluster, including Col1a1 , Col2a1 , Cxcl12 , and Tnc . Cluster 1: Col1a1 + osteoblast, Cluster 2: Col1a1 + and Col2a1 + OCT cells, Cluster 0: Cxcl12 + Adipo-CAR cell, Cluster 3: Tnc + Osteo-CAR cell. Violet: high expression, yellow: low expression. C Sub-clustering analysis of tdTomato + cells ( n = 3132 cells) extracted from the original UMAP plots. Left panels: Sub-clustering strategy for tdTomato + cells (upper). UMAP visualization of sub-clustered tdTomato + cells (lower). Upper right panel: Percentage of Dlx5 + and Fgfr3 + lineage cell distributions among each cluster. Lower right panels: Split-dot-based visualization of representative gene expression in Cluster1 (C1, osteoblasts), Cluster3 (C3, Osteo-CAR cells), and Cluster0 (C0, Adipo-CAR cells) in Dlx5 CE and Fgfr3 CE groups.
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A – C Integrating scRNA-seq analyses of Dlx5 + lineage cells, Fgfr3 + lineage cells, Col1a1(2.3 kb)-GFP-marked osteoblasts, Cxcl12-GFP-marked CAR cells at P49 (pulsed at P21). A Diagram for LIGER data integration. scRNA-seq datasets of sorted GFP + and/or tdTomato + single cells harvested from Col1a1(2.3 kb)-GFP ; Dlx5-creER ; R26R tdTomato femur bone marrow at P21 (2920 cells) and Cxcl12-GFP ; Fgfr3-creER ; R26R tdTomato (4347 cells) were merged by LIGER. Scale bar: 500 μm (whole images) or 20 μm (magnified view). B Left panel: UMAP plot of major classes of GFP + and/or tdTomato + cells (Cluster 0-12, 7267 cells). Center panels: Density plots of Col1a1(2.3 kb)-GFP ; Dlx5-creER ; R26R tdTomato (upper) and Cxcl12-GFP ; Fgfr3-creER ; R26R tdTomato derived cells (lower). Right panels: Feature plots of representative genes enriched in each cluster, including Col1a1 , Col2a1 , Cxcl12 , and Tnc . Cluster 1: Col1a1 + osteoblast, Cluster 2: Col1a1 + and Col2a1 + OCT cells, Cluster 0: Cxcl12 + Adipo-CAR cell, Cluster 3: Tnc + Osteo-CAR cell. Violet: high expression, yellow: low expression. C Sub-clustering analysis of tdTomato + cells ( n = 3132 cells) extracted from the original UMAP plots. Left panels: Sub-clustering strategy for tdTomato + cells (upper). UMAP visualization of sub-clustered tdTomato + cells (lower). Upper right panel: Percentage of Dlx5 + and Fgfr3 + lineage cell distributions among each cluster. Lower right panels: Split-dot-based visualization of representative gene expression in Cluster1 (C1, osteoblasts), Cluster3 (C3, Osteo-CAR cells), and Cluster0 (C0, Adipo-CAR cells) in Dlx5 CE and Fgfr3 CE groups.

Journal: Communications Biology

Article Title: Bone marrow endosteum houses Hedgehog-susceptible Dlx5-expressing osteoblast precursor cells

doi: 10.1038/s42003-026-09649-0

Figure Lengend Snippet: A – C Integrating scRNA-seq analyses of Dlx5 + lineage cells, Fgfr3 + lineage cells, Col1a1(2.3 kb)-GFP-marked osteoblasts, Cxcl12-GFP-marked CAR cells at P49 (pulsed at P21). A Diagram for LIGER data integration. scRNA-seq datasets of sorted GFP + and/or tdTomato + single cells harvested from Col1a1(2.3 kb)-GFP ; Dlx5-creER ; R26R tdTomato femur bone marrow at P21 (2920 cells) and Cxcl12-GFP ; Fgfr3-creER ; R26R tdTomato (4347 cells) were merged by LIGER. Scale bar: 500 μm (whole images) or 20 μm (magnified view). B Left panel: UMAP plot of major classes of GFP + and/or tdTomato + cells (Cluster 0-12, 7267 cells). Center panels: Density plots of Col1a1(2.3 kb)-GFP ; Dlx5-creER ; R26R tdTomato (upper) and Cxcl12-GFP ; Fgfr3-creER ; R26R tdTomato derived cells (lower). Right panels: Feature plots of representative genes enriched in each cluster, including Col1a1 , Col2a1 , Cxcl12 , and Tnc . Cluster 1: Col1a1 + osteoblast, Cluster 2: Col1a1 + and Col2a1 + OCT cells, Cluster 0: Cxcl12 + Adipo-CAR cell, Cluster 3: Tnc + Osteo-CAR cell. Violet: high expression, yellow: low expression. C Sub-clustering analysis of tdTomato + cells ( n = 3132 cells) extracted from the original UMAP plots. Left panels: Sub-clustering strategy for tdTomato + cells (upper). UMAP visualization of sub-clustered tdTomato + cells (lower). Upper right panel: Percentage of Dlx5 + and Fgfr3 + lineage cell distributions among each cluster. Lower right panels: Split-dot-based visualization of representative gene expression in Cluster1 (C1, osteoblasts), Cluster3 (C3, Osteo-CAR cells), and Cluster0 (C0, Adipo-CAR cells) in Dlx5 CE and Fgfr3 CE groups.

Article Snippet: Fgfr3-creER (JAX025809), Dlx5-creER (JAX010705), Rosa26-CAG-loxP-stop-loxP-tdTomato (Ai14: R26R-tdTomato , JAX007914), Col1a1(2.3 kb)-GFP (JAX013134), Ptch1-floxed (JAX030494), Rosa26-loxP-stop-loxP- iSmoM2 ( R26R- iSmoM2 , JAX005130), Rosa26-SA-loxP-GFP-stop-loxP-DTA (JAX006331), FVB/NJ (JAX001800), and C57BL/6J (JAX000664) mice were acquired from the Jackson laboratory.

Techniques: Derivative Assay, Expressing, Gene Expression