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. 2016 Oct 15;30(20):2297–2309. doi: 10.1101/gad.290619.116

Figure 6.

Figure 6.

MEF2 binding is a shared feature of many angiogenesis enhancers. (A,B) MEF2C-binding peaks are enriched around 200 kb of genes associated with sprouting angiogenesis, as assessed by increased expression in the hypersprouting retina of Dll4+/− mice (P = 8.682 × 10−09) (del Toro et al. 2010) (A) or identified through laser capture microdissection of retinal tip cells (P = 0.007776) (Strasser et al. 2010) (B). (C) Genomic snapshots denoting MEF2C-binding sites within the loci for HLX, ETS1, and ELK3 transcription factors. H3K27ac peaks are indicated in green, MEF2C-binding peaks are indicated in red, and novel angiogenic enhancers are indicated in black. (D) tol2-mediated mosaic zebrafish transgenic for the human MEF2C-binding peak enhancers HLX-3, ETS1+194, and ELK3-29 demonstrate enhancer-driven GFP expression in sprouting endothelial cells (green). (ISV) Intersegmental vessels. (E) The stable transgenic zebrafish line tg(hlx-3:GFP) directs expression specifically to sprouting endothelial cells in the intersegmental vessels. (F) Summary of reporter gene expression detected in 32-hpf tol2-mediated mosaic transient transgenic zebrafish embryos. (G) Whole-mount embryos and hindbrains of representative E11 X-gal-stained transient transgenic embryos expressing HLX-3 wild-type:LacZ and HLX-3mutMEF:LacZ transgenes. See also Supplemental Figures 6–8.