Figure 1.
Targeted disruption and embryonic expression of the Edg1 gene. (a) Schematic representation of the Edg1 targeting strategy. The structure of the mouse Edg1 locus is shown at the top, the structure of the Edg1 targeting vector in the middle, and the predicted structure of the homologous recombined locus on the bottom. RT-5′ and RT-3′, primers for RT-PCR. B, BamHI; Bgl, BglII; PBSK, pBluescript vector. (b) Genotyping of mouse offspring from the Edg1 heterozygous mating. Wild-type Edg1 locus yielded a 9.5-kb BamHI band. Disrupted Edg1 locus yielded a 2.5-kb BamHI band. No Edg1–/– mice were found born alive. (c) RT-PCR analysis of total RNA from E12.5 mouse embryos by using RT-5′ and RT-3′. Edg1+/+ and Edg1+/– RNA yielded the predicted 630-bp amplification product. No amplification product was detected from Edg1–/– RNA. (d, e) Whole-mount of Edg1+/– E9.5 and E10.5 embryos stained with X-Gal. H, heart; DA, dorsal aorta; ISA, intersomatic arteries; CP, capillaries; TC, telencephalon; ACV, anterior cardinal vein. (f) Longitudinal section of dorsal aorta (DA) from E10.5 Edg1+/– embryo. LacZ staining is seen in arterial ECs (AEC). (g) Longitudinal section of posterior cardinal vein (PCV) from E10.5 Edg1+/– embryo. LacZ staining is seen in arterial endothelial cells (AEC) but not in venous endothelial cells (VEC). (h) Transverse section of dorsal aorta from E12.5 Edg1+/– embryo. Vascular ECs and VSMCs are stained. EC, endothelial cell; VSMC, vascular smooth muscle cell. Scale bars = 50 μm.