Fig. 2

Tracing the contribution of definitive hematopoietic waves to macrophages in the embryo and early larvae (A) Scheme of the 4-OHT-inducible transgenic lines used to assess the contribution of definitive hematopoietic waves to the macrophage lineage. (B) Fluorescent images of EtOH non-switched controls (left) and 4-OHT-induced (right) Tg(fli1a:creERT2;mpeg1:Switch) embryos and larvae (2–8 dpf). Macrophages were considered as peripheral macrophages (white area) or CHT-resident macrophages (blue area). Scale bars, 100 μm. Quantification of macrophages in the periphery and the CHT are shown in (D) and (E) respectively. (C) Fluorescent images of 4-OHT-induced Tg(fli1a:creERT2;mpeg1:Switch) (16 dpf) in the periphery. Scale bar, 500 μm. Non-switched primitive macrophages (left) and definitive macrophages (right). Quantification of macrophages in the periphery is shown in (D). (D) Quantification of DsRed and GFP macrophage number in the periphery was measured over a time course of 2–16 dpf. (2 dpf n = 6; 4 dpf n = 11; 6 dpf n = 7; 8 dpf n = 4; 10 dpf n = 5 and 16 dpf n = 5). Mean ± SEM of the DsRed+ and GFP+ macrophage number at each time point is shown. two-way ANOVA with Sidak’s multiple comparison was used for this analysis. ∗∗∗∗p ≤ 0.0001. (E) Quantification of DsRed and GFP macrophage number in the CHT was measured over a time course of 2–8 dpf. (2 dpf n = 6; 4 dpf n = 11; 6 dpf n = 7; and 8 dpf n = 4). Mean ± SEM of the DsRed+ and GFP+ macrophage number at each time point is shown. Two-way ANOVA with Sidak’s multiple comparison was used for this analysis. ∗∗∗∗p ≤ 0.0001.