Extended Data Fig. 9: Splenic hematopoiesis after MI is curbed by B-cell derived acetylcholine.
From: B lymphocyte-derived acetylcholine limits steady-state and emergency hematopoiesis
a. Cd19CreChatfl/fl and Chatfl/fl mice were subjected to myocardial infarction (MI) and harvested 6 days later. HSPC in the spleen 6d after MI measured by c.f.u. assay. P = 0.0325. n = 5 mice per group, two-tailed Student’s t-test. b. Flow cytometry plots for splenic HSPC 6d after MI. c. Splenic HSPC enumeration 6d after MI. P = 0.047 for SLAM LSK, P = 0.0140 for LSK, P = 0.020 for CMP and P = 0.011 for GMP. n = 5 mice per group, two-tailed Student’s t-test. d. Leukocytes in spleen 6d after MI. n = 5 mice per group, two-tailed Student’s t-test. e. Cd19CreChatfl/fl and Chatfl/fl mice underwent splenectomy. 3 weeks after surgery, mice were subjected to MI and harvested 3 days later for flow cytometry. C57BL/6 mice were used as controls in the no splenectomy group. Flow cytometry plots for HSPC in the bone marrow 3 d after MI in splenectomized mice. f. Bone marrow HSPC enumeration. Two independent experiments, SLAM LSK: P = 0.030 for no splenectomy, P = 0.014 for splenectomy. CMP: P = 0.0095 for no splenectomy, P = 0.048 for splenectomy. SLAM LSK: n = 6 no splenectomy Chatfl/fl mice, n = 5 no splenectomy Cd19CreChatfl/fl mice, n = 8 splenectomy Chatfl/fl mice, n = 7 splenectomy Cd19CreChatfl/fl mice. CMP: n = 7 no splenectomy Chatfl/fl mice, n = 6 no splenectomy Cd19CreChatfl/fl mice, n = 8 splenectomy Chatfl/fl mice, n = 7 splenectomy Cd19Cre Chatfl/fl mice, two-tailed Student’s t-test. g. Myeloid cells and neutrophils in circulation 3 d after MI. Myeloid cells: P = 0.0053 for no splenectomy, P = 0.0009 for splenectomy. Neutrophils: P = 0.011 for no splenectomy, P = 0.016 for splenectomy. Two independent experiments, n = 12 no splenectomy Chatfl/fl mice, n = 10 no splenectomy Cd19CreChatfl/fl mice, n = 8 splenectomy Chatfl/fl mice, n = 7 no splenectomy Cd19CreChatfl/fl mice, two-tailed Student’s t-test. h. Flow cytometry plots of myeloid cells in the infarct myocardium 3 d after MI in splenectomized mice. i. Myeloid cell and neutrophil numbers in the infarct myocardium 3 d after MI. Myeloid cells: P = 0.037 for no splenectomy, P = 0.027 for splenectomy. Neutrophils: P = 0.0006 for no splenectomy, P = 0.041 for splenectomy. Two independent experiments, n = 12 no splenectomy Chatfl/fl mice, n = 9 no splenectomy Cd19CreChatfl/fl mice, n = 8 splenectomy Chatfl/fl mice, n = 7 no splenectomy Cd19CreChatfl/fl mice, two-tailed Student’s t-test. Data are mean ± SEM with individual values for data distribution, *P < 0.05, **P < 0.01 and ***P < 0.001.
