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Fig. 3 Ontogenetically distinct macrophages exhibit different migration behavior and recruitment after tissue injury during early developmental stages (A) Diagram showing the amputation plan, tail fin of Tg(fli1a:creERT2;mpeg1:Switch) were amputated at 2 dpf and macrophages recruitment to the site of injury was analyzed. Representative images are shown at 24 hpa. Scale bar: 100μm. (B) Diagram showing the counting region in the larvae. Quantification of DsRed and GFP macrophage number at the site of injury at 24 hpa (n = 14) and 48 hpa (n = 12). Mean ± SEM of the DsRed+ and GFP+ macrophage number at each time point is shown. ∗∗∗∗p ≤ 0.0001. (C) Diagram showing the amputation plan, tail fin of Tg(fli1a:creERT2;mpeg1:Switch) were amputated at 5 dpf and macrophages recruitment to the site of injury was analyzed. Representative images are shown at 24 hpa. Scale bar: 100μm. (D) Tail fin of Tg(fli1a:creERT2;mpeg1:Switch) were amputated at 5 dpf and macrophages recruitment to the site of injury was analyzed. Representative images are shown at 3 hpa. White arrows indicate the primitive macrophages with big vacuoles. Scale bar: 10μm. (E) Quantification of DsRed and GFP macrophage number at the site of injury at 3 hpa (n = 13); 6 hpa (n = 6); 12 hpa (n = 6); 24 hpa (n = 10); and 48 hpa (n = 6). Mean ± SEM of the DsRed+ and GFP+ macrophage number at each time point is shown. ns, p > 0.05; ∗∗p ≤ 0.01; ∗∗∗∗p ≤ 0.0001. (F) Quantification of dsRed and GFP macrophage number at the site of injury at 3 hpa (n = 13); 6 hpa (n = 6); 12 hpa (n = 6); 24 hpa (n = 10) and 48 hpa (n = 6). Mean ± SEM of the DsRed+ and GFP+ macrophage number at each time point is shown. Quantification was normalized by the number of total macrophages in the tail of the respective larvae (the sum of peripheral, CHT and recruited macrophages of distinct origins). ns, p > 0.05; ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗∗p ≤ 0.0001.