Real-time quantitative PCR was performed using the 2× Universal SYBR Green Fast qRT-PCR Mix (Abclonal) with the following reaction conditions: 1 cycle of 95 °C for 3 min, 40 cycles of 95 °C for 5 s, and 60 °C for 30 s. All PCR primers were designed using Primer-BLAST (NCBI) (Table 2) and were synthesized by Beijing Tsingke Biotech Co., Ltd (China). Target gene expression was normalized to the expression of the GAPDH gene, and relative expression was calculated using the 2−ΔΔCt method.
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