Td 92012

Blood Pressure (BP) was measured using implanted radio-telemetry probes PAC-10 (Data Sciences International) as described previously.^{2 (link)} Briefly, mice were anesthetized with isoflurane, 5% for initial induction and 1% to 1.5% for maintenance during surgery. The left carotid artery was isolated and the probe catheter inserted to a depth of ≈1 cm and secured with sutures. The probe body was placed subcutaneously on the left side of the abdomen. Two weeks post-surgery, continuous recordings were commenced using the Acquisition module of the Dataquest software (Data Sciences International, United Kingdom) at a sampling rate of 2 kHz for 24 to 48 hours and analyzed using Ponemah P3 plus analysis software (Data Sciences International, United Kingdom). Mice were further interrogated with a high-salt diet (8% NaCI, Envigo TD92012) and L-NAME in the drinking water (5 mg/10 mL) for 4 weeks. Twenty-four hour recordings were taken at 0, 7, 14, 21, and 28 days. At the end of the study, mice were sacrificed and the hearts harvested and weighed.
See Data Supplement for details on genotyping, organ bath and wire myography experiments, tail cuff BP measurements, analysis of atherosclerotic plaque formation, ELISA, and data analysis.

All animal protocols conform to the National Institute of Health's Guide for the Care and Use of Laboratory Animals and were approved by the University of Pittsburgh IACUC. Male C57Bl/6 mice (Jackson Laboratories) were housed in a temperature‐controlled facility on a 12 h light/dark cycle. They were kept on standard laboratory chow, a 10% KCl diet (Envigo TD.09075) for 4 days, an 8% NaCl diet (Envigo TD.92012) for 4 weeks, or given aldosterone at a rate of 240 μg/kg/day via a subcutaneous minipump (DURECT Corporation, model 2002) for 2 weeks before sacrifice. Kidneys were collected and flash frozen or fixed in 4% paraformaldehyde for 24 h before being embedded in OCT.