Extended Data Fig. 8: Contribution of neutrophils to the effects of sleep and exercise in Ldlr−/−Jak2V617FBM mice.
From: Mutation-dependent responses to sleep and exercise in clonal haematopoiesis
a. UMAP and cluster proportions of reclustered aortic CD45.2 neutrophils across lifestyle groups in Ldlr−/−Jak2V617FBM mice. Volcano plots of DEGs comparing sedentary and WT, sedentary and SF, and sedentary and exercise CD45.2 neutrophils. b. NETosis score in CD45.2 neutrophils across lifestyle groups (Elane, Mpo, Ltf, Ngp, Camp, Padi4, H2afx, Hmgb1, Ncf1, Ncf2, Ncf4, Cybb, Gsdmd, Rac2, Plcg2, Fpr1, Tlr4, Cd177). n = 5 pooled mice. c. Representative flow cytometry plots of MPO and CitH3 positivity in CD45.1 WT and CD45.2 Jak2V617F neutrophils exposed to either no stimulation, PMA/Ionomycin, or LPS. Quantification of MPO+CitH3+ CD45.2 WT or Jak2V617F neutrophils exposed to either no stimulation, PMA/Ionomycin, or LPS across the 4 lifestyle groups (n = 3 wt, 4 sed, 4 SF and 4 exercise for all three interventions and both CD45.1 and CD45.2 neutrophils, respectively. d. Experimental schematic; exemplary flow cytometry plots showing neutrophil depletion in the blood in the anti-Ly6G group compared to IgG injected controls. Bodyweight, plasma cholesterol, and plasma triglyceride levels in Ldlr−/−Jak2V617FBM mice injected with IgG or anti-Ly6G across lifestyle groups at the time of euthanasia. (n = 7 sedentary + IgG, 8 sedentary + anti-Ly6G, 7 SF + IgG, 7 SF + anti-Ly6G, 4 exercise + IgG and 7 exercise + anti-Ly6G). e. Expansion of blood CD45.2 Jak2V617F monocytes in mice injected with IgG or anti-Ly6G across lifestyle groups relative to baseline (n = 3 sedentary + IgG, 8 sedentary + anti-Ly6G, 4 SF + IgG, 8 SF + anti-Ly6G, 4 exercise + IgG and 6 exercise + anti-Ly6G). Schematics in panels c–e created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). f. Lesion area, volume, and proportion of the necrotic core in Ldlr−/−Jak2V617FBM mice exposed to sedentary lifestyle, SF, exercise plus either IgG or anti-Ly6G with representative aortic root images stained with Oil-red-O (n = 6 sedentary + IgG, 8 sedentary + anti-Ly6G, 7 SF + IgG, 7 SF + anti-Ly6G, 4 exercise + IgG and 5-6 exercise + anti-Ly6G). Scale bars are 200 µm. Two-sided statistics. Data are mean ± s.e.m; *p < 0.05, **p < 0.01. P-values were obtained from individual Mann–Whitney U tests.
