FIGURE 1

Co-localization of angpt1 with neurogenic markers in midline proliferative zone (shown schematically on the right). Double FISH in 2-dpf brain on 1.0 um thick optical sections using antisense RNA probes simultaneously hybridized against notch1a(A) and angpt1(B) and the overlay pattern of (A,B) is shown in (C). Double FISH in the 2-dpf brain using antisense RNA probes against dla(D) and angpt1(E), and the overlay pattern of (D,E) is shown in (F). Double FISH in 2-dpf brain using antisense RNA probes against nes(G) and angpt1(H), and the overlay pattern of (G,H) is shown in (I). Dynamic expression of angiogenic factors through the one-cell stage to 72 hpf by qPCR is shown in (J). P = 0.0428. (J’) Expression of angiogenic factors from 0 hpf to 10 hpf. Images are stacks of merged Z-slices. Tangerine stars indicate angpt1-positive cells co-localized with notch1a, dla, and nes-expressing cells. Statistical analysis of the qPCR result is shown in mean ± SD (N = 3 groups per stage, 20–30 embryos in one group) by Kruskal–Wallis test with Dunn’s multiple comparisons test. Scale bar is 20 μm.