Fig 1. Whole embryo or endothelial-specific inactivation of Rasa3 induces severe bleeding and lethality in E12.5 embryos.

A. The mouse Rasa3 gene structure (boxes denote exons, and exons in blue indicate the coding regions) with the corresponding protein domains, C2 (C2), the GAP-related domain (GRD) and the pleckstrin homology domain (PH), are represented. LoxP site insertions in the floxed (f) allele are indicated (red box). The post-recombination delta (∆) allele is represented. B. (Left) Immunodetection of Rasa3 and γ-Tubulin by Western blotting on extracts isolated from 5 E12.5 embryos from an R3^∆/+ mice intercross. (Right) Genotyping of embryos described at the left by PCR amplification of the genomic region between the LoxP sites (f) or of the delta Rasa3 allele. E2 embryo is R3^∆/∆, E1, E4 and E5 are R3^∆/+ embryos and E3 is the R3^f/f embryo (E3). Results are representative of 5 separate experiments. C. Uterine horns of a R3^f/f female at E12.5 after crossing with a R3^f/f iEC-Cre male, and ip injected with tamoxifen (center). Eight embryos were genotyped for the Cre transgene and the R3^∆ allele (left). Only embryos positive for the Cre transgene have the R3^∆ allele (E1, E2, E3 and E7) and present severe bleeding (right). Results are representative of 10 separate experiments.