Fig 1.
Depletion of the circulating monocyte and Gr1lo and F4/80hi populations following 1 week GW2580 treatment.
A. Identification of total monocyte population in mouse blood using MAFIA-GFP. B Following one week of GW2580 treatment, no difference in total circulating monocytes was observed. FACS quantification of Gr1hi (M1) and Gr1lo (M2; C,D) and F4/80hi (E,F) in total monocytes following 1 week GW2580 treatment (n = 6 and n = 4 animals per group, respectively).
Fig 2.
Depletion of the M2 cardiac macrophage populations following 1 week GW2580 treatment.
A. Representative profiles of digested hearts from vehicle and GW2580-treated animals. B. Quantification of cardiac tissue-resident macrophages 1 week post-GW2580 treatment (n = 6 animals per group). C. Quantification of CD206+ M2 macrophages within the heart, 1 week post-GW2580 treatment (n = 6 animals per group, nd:not detected). D. mRNA expression relative to GAPDH of M2 markers Arg1 and CD206 and E. M1 markers IL6, IL1β, respectively, within the heart following 1 week GW2580 treatment (n = 4 animals per group). * p<0.05, ** p<0.01.
Fig 3.
CD206+ M2 macrophage depletion reduces LV ejection fraction but not infarct size 2 weeks post MI in MAFIA mice.
A. mRNA expression relative to GAPDH of M1 markers IL6, IL1B, and (b) M2 markers Arg1 and CD206 respectively within the heart 2 weeks post MI (n = 4–6 animals per group). C. Determination of ejection fraction based on pressure-volume loop measurements vehicle or GW2580-treated MAFIA mice 2 weeks post MI. (* p<0.05. n = 11–13 animals per group). D. Upper panels: representative MAFIA heart sections after TTC staining. Middle panels: representative MAFIA heart section under fluorescent microscope allowing the detection of coloured microspheres distributed in the perfused area. Lower panels: Infarct areas are represented as white, non-perfused area-at risk (AAR) of infarction (dark red) and perfused areas (blue) are highlighted. The area at risk corresponds to the non-perfused area. E. AAR/LV ratio quantification in MAFIA mice 2 weeks post MI using coloured microspheres. F. IZ/LV ratio quantification in MAFIA mice 2weeks post MI using TTC staining and coloured microspheres (* p<0.05. n = 11–13 animals per group).
Fig 4.
Effect of CD206+ M2 macrophage depletion on collagen deposition and cell infiltration within the infarct 2 weeks post MI in MAFIA mice.
A. Representative Sirius Red staining on histological sections from MAFIA mice 2 weeks post MI. B. Quantification of collagen staining as a percentage of LV from MAFIA mice 2 weeks post MI (Scale bar: 1 mm, n = 6 animals per group, ≥8 images per animal). C. Representative hematoxilin and eosin staining on 20x infarct or remote histological section from MAFIA mice 2 weeks post MI showing an increase in inflammatory infiltrates in animals treated with GW2580. D. Quantification of nuclei number per mm2 in MAFIA remote and infarct zone in MAFIA mice 2 weeks post MI. (n = 4–6 animals per group). E. Representative images of immunofluorescent staining of CD206+ M2 macrophages, Gr1+ M1 macrophages and Ly6G+ neutrophils within the infarct zone from MAFIA mice 2 weeks post MI. Quantification of Ly6G+ neutrophil, Gr1+ M1 macrophage and CD206+ M2 macrophage infiltration within the infarct zone from MAFIA mice 2 weeks post MI (n = 4 animals per group, 10 images/animal). * p<0.05, ** p<0.01.