Extended Data Fig. 8: Analysis of Dll4-mCherry, Dll1-mCherry and Jag1-mCherry reporter mice.

a, Reverse-transcription PCR of Dll4, Dll1 and Jag1 in vascular cells (red), perivascular LEPR⁺ cells (purple) and osteoblasts (blue), normalized to Gapdh (n = 4 mice). b, Low-magnification immunofluorescence images of thymus sections from Dll4-mCherry, Dll1-mCherry and Jag1-mCherry mice. c, Representative flow cytometry, measuring mCherry fluorescence in total bone marrow of Dll4-mCherry, Dll1-mCherry and JAG1-mCherry mice. Indicated values represent percentages of the complete CD144⁺ and mCherry⁺CD144⁻ populations. Cells were pre-gated on DAPI⁻ cells. d, Representative mCherry levels in DAPI⁻CD45^lowTER119^lowCD144⁺ bone marrow endothelial cells from Dll4-mCherry (red) (n = 4), Dll1-mCherry (blue) (n = 3), Jag1-mCherry (black) (n = 4) and control (grey) (n = 5) mice. e, f, Representative immunofluorescence metaphysis and diaphysis of Dll4-mCherry (e) and Dll1-mCherry (f) bone marrow (n = 3 mice). mCherry (red) and LAMA1 (blue). g, h, Representative two-photon images of bone marrow from intact (left) or dextran-injected (right) Dll4-mCherry (g) and Dll1-mCherry (h) mice (n = 3 mice). i, Normalized counts of key differentially expressed genes from bulk RNA-seq performed on CD144⁻DLL1⁺ cells (purple) (from n = 2 mice) and CD144⁺DLL1⁺ cells (black) (from n = 2 mice). j, Representative flow cytometry histogram measuring mCherry fluorescence in NK1.1⁺ population from Dll1-mCherry (pink) and control (black) mice (n = 3 mice). The data are mean ± s.d. N.S., not significant, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, Student’s t-test, two-tailed. Data are representative of two (a, e–h, j) or three (b–d) independent experiments.

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