The GPCR modulator protein RAMP2 is essential for angiogenesis and vascular integrity
J. Clin. Invest. Yuka Ichikawa-Shindo, et al. 118:29 doi:10.1172/JCI33022 [Go to this article.]

Figure 1
RAMP2 expression during development and generation of RAMP2 knockout mice. (A) Real-time PCR analysis of gene expression during E11.5–E14.5 in WT embryos. Expression is shown relative to that at E11.5. n = 5 per time point. AM, CRLR and RAMPs were expressed during midgestation. (B) In situ hybridization of RAMP2 in WT embryos. Sections of umbilical artery, aortic arch, and lung from E12.5 WT embryos were used. Intense RAMP2 expression was detected in the vascular ECs. Scale bars: 20 μm. (C) Targeted disruption of mouse RAMP2. The genomic locus and predicted targeted locus are shown. Boxes denote exons 1–4 of RAMP2; ScaI and NheI restriction sites and loxP sites are indicated. Probes for Southern blot analysis are shown. (D–F) Southern blot analysis of mouse genomic DNA. (D) DNA was digested with NheI and probed with the 5′ probe. The 7.9-kb and 18.3-kb fragments denote floxed and WT alleles, respectively. (E) DNA was digested with ScaI and probed with the Neo probe. The 8.1-kb fragment denotes flox. (F) DNA was digested with ScaI and probed with the 3′ probe. The 8.1-kb ScaI fragment denotes flox; the 12.4-kb fragment denotes WT. The 4.9-kb fragment denotes the KO allele, generated by deletion of the loxP site using Cre recombinase.