2.6. Molecular Analysis
Total RNA was extracted from kidney tissues using the kit (SanPrep Column microRNA Miniprep Kit, BioBasic Inc, Canada) according to the manufacturer’s instructions. cDNA synthesis was performed using a reverse transcription kit (OneScript® Plus cDNA Synthesis Kit, Applied Biological Materials Inc, Canada). NRF2, NF-κB, NR4A2, Caspase-3 mRNA levels were measured by RT-qPCR using BlasTaq™ 2X qPCR MasterMix (Applied Biological Materials Inc., Canada). The primer set is listed in Table 6. Expression levels of target genes were interpreted based on Ct values obtained in real time PCR. Normalization was performed according to the β-actin housekeeping gene expression level and the gene expression levels were compared using the 2-ΔΔCt method21.