Absence of resident cardiac macrophages in Csf1rΔFIRE mice.
(A) Flow cytometry analysis of 3 month-old RankCreRosa26eYFP mice, showing single/CD45+/lin-(CD11c, Ter119, Tcrß, Nk1.1)/CD11b+ cardiac cells, eYFP expression in macrophages (CD64+/F4/80+) and percentage of eYFP+ blood monocytes, microglia and cardiac macrophages (n=3-5 each from an independent experiment). (B) Flow cytometry analysis of 3 month-old Flt3CreRosa26eYFP mice, showing macrophage expression of eYFP and percentage of eYFP+ blood monocytes, microglia and cardiac macrophages (n=4 each from an independent experiment). (C) Number of myeloid cells (CD45+/lin-/CD11b+), macrophages (CD45+/lin-/CD11b+/CD64+/F4/80+), neutrophils (CD45+/lin-/CD11b+/CD64-/F4/80-/Ly6g+) and Ly6chi monocytes (CD45+/lin-/CD11b+/CD64-/F4/80-/Ly6g-/Ly6chi) (n=6 for control and n=5 for ΔFIRE mice, each single experiments). (D) Representative immunohistological images showing macrophages (CD68+ cells in white and Hoechst in blue) in control and ΔFIRE hearts in 3 month-old mice at baseline conditions (scale bars represent 500µm). (E) Representative flow cytometry analysis of cardiac macrophages and (F) their expression of eYFP in Csf1rΔFIRE/+RankCreRosa26eYFP and Csf1rΔFIRE/ΔFIRERankCreRosa26eYFP. Either Fisher’s LSD test or unpaired t-test were performed and mean ± SD is shown.