In the week labeled -6, at the age of 6 weeks, male HanSD rats and TGR from several litters were exposed to 5/6 NX under general anesthesia (tiletamine + zolazepam, Virbac SA, Carros Cedex, France, 8 mg/kg; and xylasine, Spofa, Czech Republic, 4 mg/kg intramuscularly) as described previously [36 (link), 37 (link), 44 (link)]. After 24 hours’ recovery, animals were randomly assigned to experimental groups and were left with no treatment. Six weeks later (in the week 0), an appropriate treatment regime was initiated or rats were left further with no treatment. The following experimental groups were investigated:

Sham-operated HanSD rats + water (initial n = 8)

Sham-operated TGR + water (n = 8)

5/6 NX TGR + water (n = 26)

5/6 NX TGR + RAS blockade (n = 24)

5/6 NX TGR + RAS blockade + sEH blockade (n = 25)

The follow-up period was until week +60. In the weeks labeled 2, 4, 14, 22, 30, 38, 46, 54 and 60 and after appropriate habituation training, the animals were placed in individual metabolic cages and their 24-hour urine was collected for determination of daily albuminuria and urinary creatinine excretion. In addition, in the weeks 2, 4, 14, 22 and 60 daily urinary excretion of angiotensinogen was determined.