Kidney tissue sample collection and treatment
Kidney tissue was collected from 12 samples per group: the left kidney
was fixed in 4% paraformaldehyde, and the right kidney was stored at
-80 °C for mRNA and protein extraction. Proteins were extracted from
kidney tissues with RIPA lysis buffer (200 μl, BioSharp, China), and the
concentration of the harvested lysate was determined with a BCA protein
assay kit (Reith Biotechnology (Shanghai) Co., Ltd., China). Total RNA
was isolated from the kidney tissue by the TRIzol (Vazyme Biotech Co.,
Ltd., China) extraction method according to the manufacturer’s protocol.
Total RNA concentration was measured by a Nanodrop system (Thermo Fisher
Scientific, United States). cDNA was reverse transcribed with
PrimeScript RT Master Mix (Takara, Cat# RR036A) and subjected to qPCR
using specific primers.