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.