Td 94217

At 4 weeks of age, age-matched male SS-WT and SS-Arhgef11^−/− were weaned onto a low-salt diet (0.3% NaCl; TD7034; Harlan Teklad). Twenty-four-hour urine collections were done at 4 and 6 weeks of age on low-salt diet for the determination of proteinuria.^{13 (link)} After urine collections were performed at week 6, each group was randomly divided and either continued on low-salt diet (0.3% NaCl) or placed on elevated salt diet (2% NaCl, TD 94217, Harlan Teklad). The following groups were studied: low salt, SS-WT (n=16) and SS-Arhgef11^−/− (n=17); and high salt, SS-WT (n=18) and SS-Arhgef11^−/− (n=18). Urine collections were performed at weeks 8, 10, and 12 for the determination of proteinuria. After week 12 urine collections, a terminal blood pressure measurement was performed as done previously under isoflurane anesthesia.^{13 (link)} Serum samples were obtained from cardiac puncture to measure blood parameters as previously described.^{15 (link)} On euthanasia, kidney and heart weights were taken, followed by snap freezing half kidney in liquid nitrogen for downstream -omics analyses and formalin fixing the other half kidney for histological examination.

All animal procedures and protocols performed in this study were approved by the University of Toledo Institutional Animal Care and Use Committee. The procedure for the generation of a CRISPR/Cas9 targeted knock-in rat model has been previously described [3 (link)]. The non-founder S rat is a control derived from generating the knock-in. The nonfounder and the Spp2 knock-in rat strains were from stocks maintained at the University of Toledo animal facility. nonfounder and knock-in were not litter mates but were concomitantly bred for experimental procedures. Rats were weaned at 28 to 30 days of age and fed a low-salt diet (0.3% NaCl, TD 7034, Harlan Teklad). High-salt diet (2% NaCl, TD 94217, Harlan Teklad) was started at 6 weeks of age for all experiments.