Fig. 3

hif-1α is required for macrophage mobilization from the AGM region.

(a–f) Brightfield images of WISH for mfap4 expression in 54 hpf WT siblings and hif-1α^−/− in normoxia, and after hypoxia chamber or DMOG treatment for 6 h starting at 48 hpf; lateral views. Area A is situated outside the AGM region and area B represents the AGM region (outlined). Red bar marks approximate size of the dorsal aorta and blue bar that of the posterior cardinal vein. n=10 embryos from three different clutches. Scale bar, 100 μm. (g–j) Quantification of macrophage mobilization from the AGM based on mfap4 WISH experiments, showing the macrophage absolute number in area A, macrophage absolute number in area B, ratio of macrophage number in area A to macrophage number in area B and total macrophage number. (k) Maximal intensity projections of confocal z-stacks of Tg(kdrl:EGFP);Tg(mpeg:mCherry) WT and hif-1α^−/−embryos at 54 hpf. (l) Quantification of macrophage mobilization from the AGM of 54 hpf Tg(kdrl:EGFP);Tg(mpeg:mCherry) WT siblings and hif-1α^−/−embryos. (m) Mosaic vessels from transplantation of WT Tg(kdrl:ras-mCherry) donor cells into hif-1α^−/− Tg(kdrl:EGFP) host blastulae. (n) Quantification of macrophage mobilization from the AGM in control Tg(kdrl:EGFP);Tg(mpeg:mCherry) hif-1α^−/−and WT EC-transplanted hif-1α^−/−embryos at 54 hpf. Arrowheads point to macrophages in the AGM. Bars represent mean±s.e.m., n=10 embryos from three different clutches (*P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; NS, no significant changes observed; t-test). Scale bars, 50 μm.