Phenotyping and Tissue Collection in Rat Model

LH/MRrrcAek, LN/MRrrcAek, and LL/MRrrcAek rats were bred and maintained in an approved animal facility at the University of Iowa on a 12-h light-dark cycle and provided food and water ad libitum. Male offspring were used in this study. The rats were phenotyped and tissues collected as previously described [41 (link)]. Briefly, at 3 weeks of age the rats were weaned onto normal chow (Teklad 7913 – Harlan Teklad NIH-31 irradiated, 18% protein, 6% fat). At 15 weeks of age they were switched to a 4% NaCl diet (Teklad 7913 modified with 4% NaCl) until they were humanely euthanized with CO₂ at 18 weeks of age after an overnight fast. Tissues were collected and stored in RNAlater (Life Technologies, Grand Island, N.Y.) at −80 °C for subsequent RNA extraction.