Extended Data Fig. 5: Extended mechanistic analysis linking sleep and exercise to BM clonal dynamics.
From: Mutation-dependent responses to sleep and exercise in clonal haematopoiesis
a. t-SNE visualization of bone marrow scRNAseq after exclusion of minor clusters <100 cells; n = 5 pooled mice per group. b. Proliferation (Ccnd2, Ccnd3, Pim1, Npm1, Myh9, Dnmt1; left) and metabolism (Atp5k, mt-Nd3, Atp5md, Ndufb1-ps, Ndufa3, Ass1, Mospd3; right) feature scores in wild type CD45.1 and CD45.2 myeloid progenitors; n = 5 pooled mice per group. c. Heat map of select proliferation-associated gene expression in Jak2V617F myeloid progenitor cells from sedentary and exercised mice. d. Experimental setup and mitochondrial respiration marked by OCR in BMDMs isolated from Ldlr−/−Jak2V617FBM mice and sorted into mutant CD45.2 and non-mutant CD45.1 groups; n = 8 and 6 technical replicates for the sedentary CD45.1 and CD45.2 groups; n = 12 and 8 technical replicates for the CD45.1 and CD45.2 exercise groups. e. UMAP from bone marrow single-cell RNA sequencing data integrating myeloid CD45.2 progenitors from all four conditions, overlaid with a composite gene score for Il1r1 and Il1rap. f. Representative flow cytometry plots showing pro-IL-1β expression in various cell types from the BM of Ldlr−/− Jak2V617FBM mice. g. Experimental setup, and representative flow cytometry plots showing ki67 expression in bone marrow CMPs in WTJak2V617FBM mice injected with saline or IL-1β, as well as quantification of %ki67 positivity in CMPs. n = 3 for saline groups, n = 5 for IL-1β groups. h. Bodyweight, plasma cholesterol, and plasma triglyceride levels in Ldlr−/−Jak2V617FBM mice with and without SF, with IgG or anti-IL-1β at the time of euthanasia. n = 10, 10, 9 no SF + IgG; 7, 6, 6 no SF + anti-IL-1β; 13, 13, 13 SF + IgG and 8, 7, 7 SF + anti-IL-1β for bodyweight, cholesterol and triglycerides, respectively. i. Flow cytometry plots for ki67 in CMPs from Ldlr−/−Jak2V617FBM mice with and without SF, with IgG or anti-IL-1β. Two-sided statistics. Data are mean ± s.e.m; ns= not significant, *p < 0.05, **p < 0.01,****p < 0.0001. P-values were derived from ordinary one-way ANOVA tests. Schematics in panels d–i created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026).
