Figure 1

Figure 1

Generation of the transgenic HRE-sv40mp:GFP reporter zebrafish.A, Schematic representation of the Tol-2 vector used to generate the Tg (hre-sv40mp: GFP) zebrafish. The construct includes a 68-bp fragment from the hypoxia-response element (HRE) of the human ENO1 gene and the SV40 minimal promoter (SV40mp). B, Representative images show the Tg (hre-sv40mp:GFP) embryos at 48 and 72 hpf under normoxic or hypoxic conditions. Tg (hre-sv40mp:GFP) embryos were exposed to hypoxia (10% O₂) for 24 or 48 h from 24 hpf. Transgenic embryos show strong GFP expression during hypoxia. However, GFP expression is observed only in the visceral organs in the transgenic embryos exposed to normoxia. C, Tg(hre-sv40mp:GFP) embryos injected with the dominant-active (DA) HIF-1α mRNA at the 1-cell stage show strong GFP expression at 48 hpf. D, Transgenic zebrafish in the vhl^−/− background show strong GFP fluorescence compared to the sibling embryos at 72 hpf. Scale bar, 500 μm.