Generation and characterization of the Tie2RAGE mouse line. (A) Tie2RAGE construct. Genomic RAGE DNA was cloned between the Tie2 promoter and enhancer. Gray boxes indicate RAGE exons; Roman numerals indicate RAGE exon numbers. E, EcoRI restriction enzyme site. (B) Southern blot analysis of DNA obtained from tail biopsies from WT and Tie2RAGE mice; EcoRI digestion releases a 2.2-kb DNA fragment from the Tie2RAGE transgene. (C) RT-PCR of different tissues derived from Tie2RAGE (Tie2) and WT mice analyzed for RAGE mRNA expression. WT indicates transgene-negative littermates. HPRT was used to demonstrate equal loading. (D) Immunohistology for RAGE protein in Tie2RAGE and WT kidney tissue. Brown staining indicates RAGE expression; magnification, ∞400. RAGE expression in kidneys of Tie2RAGE mice is evident in the endothelium of large and small vessels, while no RAGE protein could be detected in kidneys of healthy WT mice.