*2.3. Quantitative Real-Time PCR*

Total cellular RNA was extracted using a total RNA rapid extraction kit (Fastagen, Shanghai, China) according to the instructions, and 1 μg RNA of each sample was subsequently reverse transcribed to cDNA with a reverse transcription kit (TaKaRa, Dalian, China) according to the manuals. The acquired cDNA was then used as the template in a qPCR assay by using TB Green® Premix Ex Taq™ II (Tli RNaseH Plus) or Premix Ex Taq™ (Probe q-PCR)(Takara Biomedical Technology, Beijing) in CFX96 Real-time polymerase chain reaction system (qPCR) (Bio-Rad, CA, USA). The abundance of individual gene mRNA transcripts in each sample was measured three times, and GAPDH mRNA was used as the endogenous loading control. The sequences of primers and probe are listed in Table 1. Relative mRNA expression of each target gene was calculated by the 2−ΔΔCT method.


**Table 1.** Sequences of the primers and probe used for Real-time polymerase chain reaction (PCR).


**Table 1.** *Cont*.

F: forward primer; R: reverse primer.
