Fig. 4

Depletion of both notch2 and notch3 hardly affected Notch-Rbpj-mediated transcriptional activity in ECs. (A) Confocal stack images of the trunk vasculature of the wild-type (WT) sibling or notch2/notch3 mutants with the Tg(fli1a:H2B-GFP);Tg(TP1:H2B-mCherry) background at 74?hpf. (B) The percentage of ECs in observed trunk region with TP1:H2B-mCherry expression, as observed in A. Total number of EC nuclei marked by fli1a:H2B-GFP and the number of EC nuclei positive for both fli1a:H2B-GFP and TP1:H2B-mCherry were counted using Imaris software. Bars and circles indicate averages and each value of observed larvae, respectively. (C) Confocal images of trunk vasculature of the 50?hpf Tg(fli1a:H2B-GFP);Tg(TP1:H2B-mCherry) embryos injected with 10?ng control MO (left), 5?ng each of notch2 and notch3 MOs (middle, N2/N3 MOs) or 10?ng of dll4 MO (right). (D) The expression of deltaC or ephrin-B2a mRNA, as detected by whole-mount in situ hybridization, in 48?hpf embryos injected with 10?ng control MO or 5?ng each of notch2 and notch3 MOs. Numbers indicate embryos with the similar expression as shown in each image/total number of embryos. Two independent experiments were performed for each marker with at least seven embryos per experiment. In A and C, arrowheads indicate ECs with TP1:H2B-mCherry expression; boxed regions are enlarged to the right. Scale bars: 50?µm; 10?µm in enlarged images. DLAV, dorsal longitudinal anastomotic vessels; PCV, posterior cardinal vein.