Wild type sprague dawley rats

All procedures involving animals were conducted according to the UCL Ethics Committee guidelines and the UK Animals Scientific Procedures Act UK (1986) and its Amendment Regulations (2012). Wildtype Sprague-Dawley rats (Charles River, Wilmington, USA) were used in this study.

For qRT-PCR experiments, wild-type Sprague-Dawley rats were obtained from Charles River. The mutant mouse line Dst^dt-27J was used for all other experiments. This line arose from a spontaneous mutation in the Dst allele, which confers phenotype in a recessive manner. The mutant animals were first identified at the Jackson Laboratory, and were then characterized as expressing very low levels of neuronal Dst transcript levels relative to wild-type animals [27 (link)]. The animals were sacrificed at P0-P1 for primary OL cell culture and at P15 for in vivo analyses. Genotypes were determined by PCR amplification of genomic tail DNA. The University of Ottawa Animal Care Committee approved all experimental protocols. The protocols conformed to or exceeded those defined in the Canadian Council on Animal Care's Guide to the Care and Use of Experimental Animals, and the Animals for Research Act.

Wild-type Sprague-Dawley rats (Male, 4–5 weeks, Charles River) were housed under a controlled environment with a 12 h light-dark cycle and access to food and water ad libitum. All animal experiments were performed in accordance with the guidelines established and approved by the Animal Care and Use Committee at Brigham and Women's Hospital, Harvard Medical School (2016N00146). After Wild-type Sprague-Dawley rats were acclimated for a week in the BWH CCM animal facility. A single dose of puromycin aminonucleoside (50 mg/kg, PAN group) was given i.p. to rats to induce nephrosis, and PBS was given as control. Following the PAN injection, vehicle or AC1903 (50 mg/kg) was administered twice daily (at 9 am and 9 pm) for seven days. Twenty-four-hour urine albumin levels were measured on day 0, 3, and 7. Rats were euthanized after the metabolic collection on day 8. Both kidneys were collected for downstream experiments. In most cases, one kidney was used for acute glomeruli isolation and glomerular single-channel recording. For immunostaining, another kidney was quickly removed and cut into half. One half was flash frozen in liquid nitrogen, and the other was fixed in 4% PFA overnight and stored in PBS for follow-up experiments. In combination, we have studied 34 rats (PBS group n = 6, PAN group n = 15, PAN + AC1903 group n = 13).