Figure 2: Development of GRAF3 hypomorphic mice.
From: The smooth muscle-selective RhoGAP GRAF3 is a critical regulator of vascular tone and hypertension
(a) Schematic of gene trap insertion site in the SIGTR CE0477 ES clone. 5′ RACE revealed the gene trap cassette (β-geo) was located in intron 3 (see Supplementary Table S1), and this locus was verified and further refined using a targeted PCR approach that revealed the site of integration between chromosome 9: 9,082,984–9,083,121 (reverse strand). (b) RT–PCR analysis of GRAF3 (exon 1–4) and GAPDH mRNA levels in tissues from Wt and GRAF3gt/gt mice (quantification shown graphically below; mean±s.e.m., n=3/genotype). Similar results were found using primers to target downstream exons (see Supplementary Fig. S2b). (c) GRAF3 expression pattern revealed by whole mount Lac Z staining (blue; scale bars, 1.5 mm) or by dual immunohistochemical staining for β-GAL (green; scale bar, 10 microns), PECAM (red), (DAPI, blue) in frozen sections (coronary vessel, top–middle) from GRAF3+/gt mice. Vessel-restricted LacZ staining was also evident in brain (see Supplementary Fig. S2f). No significant LacZ staining or β-GAL immunofluorescence was observed in tissues from Wt mice. Mes., mesenteric.
