Fig 2: Macrophage Rmp protects against myocardial infarction. (a) Study design of bone marrow transplantation experiments, in which the mice were induced MI. (b) Representative images of infarct size by Masson's trichrome staining 7 days after MI. (c) Quantitative analysis of fibrosis as a percentage of total myocardial tissue area. n = 4–6 per group. Values are mean ± SEM. Student's t test, **P < 0.01. (d) Representative echocardiography (M-mode) 7 days after MI. (e) Echocardiographic analysis of LVEF and LVEDV (n = 5; Student's t test, *P < 0.05 and **P < 0.01). (f) Immunohistochemical staining of collagen I accumulation at the infarct site 7 days after MI. Scale bar = 200 µm. (g) Quantitative analysis of collagen I (n = 5; Student's t test, *P < 0.05).

Fig 3: Macrophage Rmp deficiency promotes M1 polarization and inhibits angiogenesis after myocardial infarction. (a) Three-color flow cytometry shows the M1 (CD68+/iNOS+) and M2 (CD68+/CD206+) macrophages at the infarct site 7 days after myocardial infarction (MI). (b) Quantitative analysis of M1 and M2 macrophages (n = 5; Student's t test, ***P < 0.001). (c) Representative images of CD206 and iNOS by western blotting 7 days after MI. (d) Corresponding densitometric quantification of the immunoreactive bands. Values are mean ± SEM of n = 5 per group. ***P < 0.001 versus LC by unpaired Student's. (e) Two-color immunofluorescence shows CD31 in LC and MRKO mice 7 days after MI. (f) Quantitative analysis of CD31. Values are mean ± SEM of n = 5 per group. ***P < 0.001 versus LC by unpaired Student's. (g) Representative images of CD31 in the infarct area of LC and MRKO mice by western blotting 7 days after MI. (h) Quantitative analysis of CD31. Values are mean ± SEM of n = 5 per group. ***P < 0.001 versus LC by unpaired Student's.

Fig 4: Macrophage Rmp promotes macrophage M2 polarization and angiogenesis after myocardial infarction. (a) Representative images of CD206 and iNOS by western blotting 7 days after myocardial infarction (MI). (b) Corresponding densitometric quantification of the immunoreactive bands. Values are mean ± SEM of n = 4 - 6 per group. *P < 0.05, **P < 0.01, and ***P < 0.001 by unpaired Student's. (c) Two-color immunofluorescence analysis shows CD31 in mice receiving the indicated bone marrow-derived macrophages (BMDMs) 7 days after MI. (d) Quantitative analysis of CD31. Values are mean ± SEM of n = 5 per group. *P < 0.05, **P < 0.01, and ***P < 0.001 by unpaired Student's. (e) Representative image analysis of CD31 in the infarct area of mice receiving the indicated BMDMs by western blotting 7 days after MI. (f) Quantitative analysis of CD31. Values are mean ± SEM of n = 5 per group. *P < 0.05, **P < 0.01, and ***P < 0.001 by unpaired Student's.

Fig 5: Macrophage Rmp deficiency aggravates cardiac dysfunction after myocardial infarction. (a) Survival curves for macrophage-specific Rmp-knockout (MRKO) and littermate control (LC) mice. ***P < 0.001. N = 15 mice per group. (b) Representative images of the hearts from LC and MRKO mice collected 7 days after myocardial infarction (MI). Scale bar = 2 mm. (c) Representative images of infarct size after Masson's trichrome staining of the hearts collected 7 days after the induction of MI. (d) Quantification of fibrosis as a percentage of total myocardial tissue area. Values are mean ± SEM of n = 6 per group. Student's t test, *P < 0.05. (e) Representative echocardiography (M-mode) and left ventricular function 7 days after the induction of MI. (f) Echocardiographic analysis of LVEF and LVEDV (n = 5; Student's t test, ***P < 0.001). (g) Immunohistochemical staining of collagen I accumulation at the infarct site 7 days after the induction of MI. Scale bar = 200 µm. (h) Quantitative analysis of collagen I (n = 5; Student's t test, ***P < 0.001).