Fig. S2

Fig. S2

peak1 is required for vascular development in zebrafish embryos. (a) Schematic showing design of splicing morpholino to target the zebrafish peak1 gene. (b) Schematic showing the design of TALEN used to target the zebrafish peak1 gene. (c) Bright field image show normal pericardial cavity and circulation of Ctrl MO injected zebrafish embryos. Scale bar = 50 μm. (d) Bar graph represents relative peak1 mRNA level versus actn1 from 30 hpf embryos treated with Ctrl MO, SP1MO or SP2MO and analyzed by qPCR. (e) Western blot analyses for indicated proteins of 30 hpf embryos treated as in Figure 1c. (f) Genotyping of zebrafish peak1 TALEN F0 founder embryos. Primers TALEN test F+R and restriction enzyme MslI were used. Uncut (%), the percentage of uncut band out of the total DNA. (g) Bar graph represents percentage of animals with blood circulation defects caused by injection of zebrafish peak1 TALEN mRNAs. (h) Bright field images of zebrafish embryos showing severe pericardial edema (arrows) and blood circulation defects (arrowheads) caused by peak1 TALEN mRNAs at indicated dpf. Scale bar = 1 mm. (i) Confocal fluorescence images of the tail vasculature of Tg(fli1:egfp)^y1 embryos treated with peak1 TALEN mRNAs were captured at 2dpf. Arrows point to mosaic stunted and disrupted ISV vessels. Scale bar = 50 μm. (j) DNA sequencing results of F1 TALEN mutant lines. (k) peak1^{Δ2/ Δ2} TALEN homozygous mutants were incrossed and progeny embryos were lysed at 2dpf and analyzed by western blotting with indicated antibodies. All data are representative of at least three independent experiments. ***, P<0.001; *, P<0.05; N.S., not significant.