To lyse cells, 1 mL Trizol reagent (TaKaRa, Japan) was added to the sample tissue according to the actual manufacturer’s instructions and incubated on a shaker for 15 min at room temperature. Total RNA was subsequently extracted from target samples. One microgram of RNA was reverse transcribed into cDNA using the Revert Aid First Strand cDNA Synthesis Kit (Thermo, United States). Quantitative RT-PCR was then performed with Pro Taq HS Premix Probe qPCR Kit (Accurate, Hunan, China). The GAPDH gene was used as an endogenous control gene for normalizing the expression of target genes. Primers used in this study included CASP9 (forward 5′-CTGTCTACGGCACACAGATGGAT-3′, reverse 5′- GGGACTCGTCTTCAGGGGAA-3′), GAPDH (5′-CAGGAGGCATTGCTGAT-3′, 5′-GAAGGCTGGGGCTCATTT-3′).
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