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Validation of DEGs using quantitative real-time PCR analysis Quantitative real-time polymerase chain reaction (qRT-PCR) was employed using the SYBR Green PCR Master Mix (Takara, Dalian, China) following the manufacturers instructions at an ABI ViiA 7 Real-time PCR platform with a total reaction volume of 20 L, containing 10 L of SYBR GreenMaster, 2 L of diluted cDNA template, 1 L of each primer (10 M), and 7 L of water to examine the reliability of RNA-seq results