Extended Data Fig. 2: Increased stem and progenitor cell quiescence after 6 weeks of exercise. | Nature Medicine

Extended Data Fig. 2: Increased stem and progenitor cell quiescence after 6 weeks of exercise.

From: Exercise reduces inflammatory cell production and cardiovascular inflammation via instruction of hematopoietic progenitor cells

Extended Data Fig. 2

Mice were given 5-bromo-2- deoxyuridine (BrdU) intraperitoneally (1 mg). BrdU incorporation in (a) long-term hematopoietic stem cells (LT-HSC), short-term HSC (ST-HSC), (b) common myeloid progenitors (CMP, ***p = 0.00026), megakaryocyte erythroid progenitors (MEP, ***p = 1.028 × 10−5), granulocyte macrophage progenitors (GMP, ***p = 4.17 × 10−4, all n = 14 animals for sedentary, n = 15 for exercise), macrophage and dendritic cell progenitors (MDP, **p = 0.0070, n = 7 animals per group) and B cell progenitors (B cell prog, **p = 0.0065, n = 6 animals per group) were analyzed 22 hours later (two-tailed U test for CMP, MDP and B cell prog; two-tailed Student’s for MEPs and GMPs). (c) Flow cytometry gating for hematopoietic progenitors and representative flow cytometry plots of BrdU gating. (d) Cell cycle analysis in LSK assessed by Ki-67/ DAPI staining. Representative flow cytometry dot plots of + LSK (*p = 0.038, n = 7 animals for sedentary, n = 12 for exercise, 2 independent experiments, two-tailed U test). (e) Experimental outline for BrdU pulse-chase experiment. Mice received BrdU in drinking water for 3 weeks (baseline, n = 3 animals) prior to 3 weeks of exercise. (f) BrdU incorporation into LT-HSC, ST-HSC, multipotent progenitors (MPP, **p = 0.0074), CMP (p = 0.14), MEP (*p = 0.034) and GMP (p = 0.07, n = 9 animals for sedentary, n = 4 for exercise, two-tailed U test comparing sedentary and exercise). (g) Representative images of granulocyte macrophage colonies from sedentary and running mice. (h) Bone marrow unit assay (CFU) of bone marrow mononuclear cells (BMNCs) for complete colonies (*p = 0.036, n = 6 animals per group, 2 independent experiments, two-tailed U-test). (i) Number of HSPC per femur in sedentary and exercising mice (n = 15 animals per group). (j) Number of marrow leukocytes at Zeitgeber 13: B cells (**p = 0.0089), CD4 T cells, CD8 T cells (*p = 0.048), neutrophils (*p = 0.044), monocytes (*p = 0.041), eosinophils (*p = 0.019, n = 5 animals for sedentary, n = 6 for exercise) and NK cells (**p = 0.00099, n = 3 per group, two-tailed U test). (k) Numbers of platelets (*p = 0.016, two-tailed Student’s), red blood cells (RBC), hemoglobin (HGB) and hematocrit (HCT, n = 12 animals for sedentary and n = 11 for exercise, 4 independent experiments). Data are mean ± s.e.m.We acknowledge Servier Medical Art (https://smart.servier.com) for providing images of mice and components of the cartoon.

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