Extended Data Fig. 10: Acetylcholinesterase inhibitor contains hematopoiesis and inflammation after MI.

a. C57BL/6 mice receiving either saline or pyridostigmine for 4 weeks were subjected to myocardial infarction (MI) and assayed 3 days after MI. Flow cytometry plots for HSPC. b. Bone marrow HSPC, CMP, GMP and MDP enumeration using flow cytometry. P = 0.019 for SLAM LSK, P = 0.039 for CMP, P = 0.0059 for GMP and P = 0.0057 for MDP. Two independent experiments, n = 12 saline and n = 11 mice pyridostigmine, two-tailed Student’s t-test. c. C.f.u. assay of bone marrow mononuclear cells (BMNCs) for granulocytes and macrophages progenitors (GMs). P = 0.040. n = 5 saline, n = 4 mice pyridostigmine group, two-tailed Mann-Whitney test. d. Circulating HSPC measured by c.f.u. assay. P = 0.0159. n = 5 saline, n = 4 mice pyridostigmine, two-tailed Mann-Whitney test. e. Flow cytometry plots for circulating myeloid cells. f. Leukocyte subsets in circulation. P = 0.0015 for leukocytes, P = 0.026 for monocytes, P = 0.046 for Ly6^high monocytes and P = 0.026 for neutrophils. Two independent experiments, n = 13 saline, n = 11 pyridostigmine mice, two-tailed Student’s t-test. g. Flow cytometry plots of myeloid cells in the infarct myocardium 3 days after MI. h. Myeloid cell numbers in the infarct myocardium 3 days after MI. P = 0.0093 for myeloid cells, P = 0.0056 for neutrophils, P = 0.0460 for monocytes and P = 0.0140 for macrophages. Two independent experiments, n = 13 saline, n = 11 pyridostigmine mice, two-tailed Student’s t-test. i. Myeloid cell numbers in the remote myocardium 3 days after MI. P = 0.049 for macrophages. Two independent experiments, n = 13 saline, n = 11 pyridostigmine mice, two-tailed Student’s t-test. i. C57BL/6 mice receiving either saline or pyridostigmine for 4 weeks were subjected to MI and harvested 7 days after MI. Immunohistochemical staining and quantification of CD11b⁺ myeloid cells, collagen I, α-smooth muscle actin⁺ myofibroblasts and CD31⁺ vessels in the infarct border zone 7d after MI. Bar graphs show percentage of positive staining per region of interest (ROI). Each data point represents a ROI and 4 ROI were analyzed per mouse. P = 0.023 for CD11b and P = 0.0041 for collagen I. Two independent experiments, n = 4 saline, n = 3 pyridostigmine mice, two-tailed Student’s t-test. k. Survival after MI. P = 0.697. Two independent experiments, n = 9 saline, n = 10 pyridostigmine mice, Mantel–Cox log-rank test. Data are mean ± SEM with individual values for data distribution, *P < 0.05 and **P < 0.01.