*4.3. Reverse Transcription PCR and qRT-PCR*

Total RNA (1 µg) from each sample was used in reverse transcription. First-strand cDNA was synthesized with a PrimeScript™ RT reagent kit with a gDNA Eraser kit (TaKaRa, Japan). The concentration of the synthesized cDNA was determined, and the cDNA was diluted to 20 ng/µL for PCR analysis. Semi-RT-PCR was performed, which consisted of denaturation at 94 ◦C 5 min; followed by 25 cycles of 94 ◦C for 30 s, 55 ◦C for 30 s, and 72 ◦C for 60 s. The qRT-PCR conditions were pre-denaturation at 95 ◦C for 30 s; followed by 30 cycles of 95 ◦C for 5 s, 60 ◦C for 20 s, and 72 ◦C for 15 s. All primer sequences used in this study are listed in Table S4. The semi-RT-PCR products were separated on 1.5% agarose gels, and stained with ethidium bromide. The qRT-PCR results were analyzed using the 2−∆∆<sup>C</sup> <sup>T</sup> method, with three biological replicates.
