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.