cDNA encoding human AT-1 was isolated by BamHI and EcoRV digestion from Topo-AT-1 constructs and subcloned into pTRE-Tight vector (Takara Bio Inc.). TRE-AT-1 Tg lines were generated by injection of linearized pTRE-Tight vector (Takara Bio Inc.) containing AT-1 cDNA into the pronucleus of fertilized eggs from FVB mice. Monogenic pTRE-AT-1 mice were backcrossed to WT C57BL/6 mice for five generations, and then bred to CamK2a-tTA mice (B6.Cg-Tg(Camk2a-tTA)1Mmay/DboJ; The Jackson Laboratory), generating nonTg WT, Camk2a-tTA monogenic, TRE-AT-1 monogenic, and CamK2a-tTA;TRE-AT-1 (referred to as AT-1 Tg) mice. Genotyping from tail DNA was performed using the following primers: AT-1 forward (5′-AATCTGGGAAACTGGCCTTCT-3′), AT-1 reverse (5′-TATTACCGCCTTTGAGTGAGCTGA-3′), Camk2a-tTA forward (5′-CGCTGTGGGGCATTTTACTTTAG-3′), and Camk2a-tTA reverse (5′-CATGTCCAGATCGAAATCGTC-3′).
Animal experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and were approved by the Institutional Animal Care and Use Committee of the University of Wisconsin-Madison and the Madison Veterans Administration Hospital. Unless specified, WT littermates were used as control throughout our study.
Animal experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and were approved by the Institutional Animal Care and Use Committee of the University of Wisconsin-Madison and the Madison Veterans Administration Hospital. Unless specified, WT littermates were used as control throughout our study.