Quantitative real-time PCR analysis
Total RNA was extracted from kidney tissue using total RNA extraction
Kit (BioTek, Winooski, VT, United States). Reverse transcription was
performed using PrimeScript RT Reagent Kit (Takara Bio, Inc., Otsu,
Japan) according to instruction. Reactions of PCR amplification were
quantified using the iTaq Universal SYBR Green Supermix (Bio-Rad
Laboratories, Inc.) in a PCR system (CFX Connect; Bio-Rad, Hercules, CA,
United States). Results were presented as relative mRNA levels to GAPDH.