Fig. 3: Cell-fate analysis reveals dormancy of Cxcl12-creER+ BMSCs. | Nature Communications

Fig. 3: Cell-fate analysis reveals dormancy of Cxcl12-creER+ BMSCs.

From: A Wnt-mediated transformation of the bone marrow stromal cell identity orchestrates skeletal regeneration

Fig. 3: Cell-fate analysis reveals dormancy of Cxcl12-creER+ BMSCs.The alternative text for this image may have been generated using AI.

a–l Long-chase analysis of Cxcl12-creER+ cells. a–c Col1(2.3 kb)-GFP; Cxcl12-creER; R26RtdTomato distal femurs with growth plates on top, analyzed at 1Y. Pulsed at P3 (a), P21 (b), and 8 W (c). Yellow double arrows: tdTomatoneg area beneath growth plates, white brackets: tdTomato+ domain in marrow space. Scale bar: 500 µm. Gray: DIC. d Quantification of tdTomato+ reticular cells in marrow space at 1Y, based on distance from growth plate. Pulsed at P3 (blue line), P21 (green line) or 8 W (red line). n = 6 (P3, P21-pulsed), n = 5 (8W-pulsed) mice. e Cxcl12GFP/+; Cxcl12-creER; R26RtdTomato bone marrow at 1 y (pulsed at P21), central marrow space. Gray: DAPI. Scale bar: 20 µm. n = 3 mice. f Quantification of tdTomato+ reticular cells in marrow space during extended chase, pulsed at 8 W. n = 4 (9 W), n = 5 (3 M), n = 3 (6 M, 1 y, and 1 y 6 M) mice. g Col1(2.3 kb)-GFP; Cxcl12-creER; R26RtdTomato bone marrow at 1 y (pulsed at P21), endosteal space. Scale bar: 20 µm. n = 4 mice. h Percentage of Col1(2.3 kb)-GFP+tdTomato+ cells per total Col1(2.3 kb)-GFP+ cortical bone osteoblasts. (Left): flow cytometry. n = 4 (+1 W), n = 3 (+8 W) mice. (Right): labeled at P3, P21, 8 W, and 5 M, analyzed at 1 y 6 M. OB osteoblasts, OCY osteocytes. n = 3 per group. i Col1(2.3 kb)-GFP; Cxcl12-creER; R26RtdTomato distal femurs at 9 W (pulsed at 8 W). 1 (endosteal area): boxed area on left panel. Gray: DIC. Scale bar: 500 µm (left), 20 µm (right). n = 3 mice. j Cxcl12-creER; R26RtdTomato distal femur bone marrow at 1Y (pulsed at 8 W). Lower left panel: central marrow space with Cxcl12GFP/+, right panels: endosteal space and trabecular bone with Col1(2.3 kb)-GFP. Gray: DIC. Scale bar: 20 µm. n = 3 mice. k, l Percentage of Col1(2.3 kb)-GFP+tdTomato+ cells per total Col1(2.3 kb)-GFP+ cells by using flow cytometry (k). n = 3 (+2 days), n = 4 (+1 W), n = 7 (+4 W) and n = 6 (+8 W) mice. l Labeled at 8 W, analyzed at 9 W, 3 M, 6 M, 1 y, and 1 y 6 M. n = 3 (9 W, 3 M, 1 y, and 1 y 6 M), n = 4 (6 M) mice. ****p < 0.0001, ***p < 0.001, two tailed, one-way ANOVA followed by Tukey’s post hoc test (f, h, k, l). Two-tailed, Mann–Whitney’s U test (h). All data are presented as mean ± s.d. Source data are provided as a Source Data file.

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