Fig. 1

peak1 is required for vascular development in zebrafish embryos.

a Bright-field images of pericardial edema (upper panel, arrowhead) and blood accumulation in the anterior aorta as well as in the tail region (arrows) of peak1 morpholino (MO) knockdown embryos (SP1MO). Pictures were taken at 2 days-post-fertilization (2dpf). Also see Supplementary Fig.S2c. b Bar graph shows the percentage of animals with vascular defects caused by control (Ctrl MO) or two different peak1 specific morpholinos SP1MO and SP2MO. Mean ± SEM; n = 30. c Confocal images at indicated time points of vasculature formation of Tg(fli1:egfp)^y1 zebrafish embryos treated as in (a). ISVs = intersegmental vessels. DLAVs = dorsal longitudinal anastomotic vessels. DA = dorsal aorta. PCV = posterior cardinal vein. hpf = hours-post-fertilization. Arrows point to disorganized endothelial sprouts and vessel structures. d Bar graph depicts the average ISV length at 32 hpf of MO-treated embryos. Mean ± SEM; n = 50. e Confocal images of subintestinal vessels (SIVs, dashed line circled region) were captured at 3dpf from embryos treated as in (c). Arrows show SIV defects. f Bar graph shows the cumulative length of SIVs per embryo from embryos treated as in (e); Mean ± SEM; n = 10. g Bar graph represents the average ISV length at 32hpf of embryos treated as in (c) and co-treated with peak1 mRNA or a nonspecific control mRNA (Ctrl mRNA). Mean ± SEM; n = 50. h Bar graph shows the percentage of Tg(fli1:egfp)^y1embryos with mosaic disrupted ISV defects after treatment of peak1 specific transcription activator-like effector nuclease (TALEN) mRNAs. TALEN R = right arm (120 pg); TALEN L = left arm (120 pg); TALEN R + L = both arms (60 pg each); Mean ± SEM; n = 90. i Embryos from incrossed heterozygous peak1 ^{∆2/ +} zebrafish were injected with SP1MO at one-cell stage, subsequently selected at 72 hpf for the vascular defects (Sensitive to SP1MO) or randomly selected and all genotyped individually. Bar graph represents percentage of homozygous peak1^{∆2/ ∆2} in each group. Mean ± SEM; n = 90. All the data are representative of at least three independent experiments. ***, P < 0.001; **, P < 0.01; N.S., not significant; in (b, d), vs. Ctrl MO; in (h), vs. TALEN R + L. Scale bar = 50 μm