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. 2022 Oct 27;10:62. doi: 10.1038/s41413-022-00232-x

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© The Author(s) 2022, corrected publication 2022

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Fig. 4 — A suppressive immune response is a hallmark of HO propagation with bone loss. a Statistical analysis of peripheral WBCs, lymphocytes and monocytes in the WT and NSE-BMP4 mice at 7 dpi (n = 5 from three independent experiments). Data are presented as the mean ± s.d. of biological replicates. ^*P < 0.05, ^**P < 0.01, N.S. indicates no significance (unpaired two-tailed t test). Statistical analysis of the expression of Cd206 (b) and Cd4 (c) in the WT and NSE-BMP4 mouse muscle via an RNA-sequencing assay (n = 3 per group). Data are presented as the mean ± s.d. of biological replicates. ^*P < 0.05, ^**P < 0.01. N.S. indicates no significance. d Representative flow cytometry images of macrophage polarization in uninjured or injured muscle from the WT and NSE-BMP4 mice. Statistical analysis of the population of M1 macrophages (e) and M2 macrophages (f) in uninjured or injured muscle from the WT and NSE-BMP4 mice (n = 3 per group). Data are presented as the mean ± s.d. of biological replicates. ^*P < 0.05, ^***P < 0.001. N.S. indicates no significance (unpaired two-tailed t test). g–j, Statistical analysis of the expression of Cd27, Cd40, Pdcd1 and Havcr2 in injured muscle from the WT and NSE-BMP4 mice at different postinjury times (n = 3 per group). Data are presented as the mean ± s.d. of biological replicates. ^*P < 0.05, ^**P < 0.01. N.S. indicates no significance (unpaired two-tailed t test). k At 7 dpi, immunostaining showed colocalization of CD206 or CD8 and immune checkpoint molecules in HO lesions. l, m Statistical analysis of the population of CD206⁺ or CD8⁺ cells expressing immune checkpoint molecules (n = 3 per group, n = 3 ROI/mouse). Data are presented as the mean ± s.d. of biological replicates. ^**P < 0.01, ^***P < 0.001, ^****P < 0.000 1. N.S. indicates no significance (unpaired two-tailed t test)