Then cDNA was synthesized following the manufacturer’s protocol of PrimeScript™ II 1st strand cDNA synthesis kit (Takara Bio, Beijing, China). Primers were designed using Primer Premier 5.0 (Premier Biosoft International, San Francisco, CA, USA) and synthesized by Qingke Biotech (Guangzhou, China). Detailed primer sequences are listed in Table S4. A 10 μL reaction mixture, containing 5 μL of 2× TB Green Premix Ex Taq II Fast qPCR Mix (Takara Bio, Beijing, China), 0.5 μL of each forward and reverse primer, 0.2 μL of cDNA template and 3.8 μL of RNase-free water, was carried out following the thermal cycling conditions: 95 °C for 3 min; 39 cycles of 95 °C for 10 s, annealing temperature (Tm) for 10 s, 72 °C for 10 s; followed by a melt curve analysis.
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