Real-time quantitative PCR

Total RNA was extracted using TRIzol reagent (Invitrogen, USA) according to the manufacturer’s instructions. RNA concentration and purity were assessed using a NanoDrop 2000 Spectrophotometer (Thermo Scientific, USA). In total, 2 mg of RNA was reverse transcribed using the high-capacity cDNA Reverse Transcriptase (RT) Kit (Invitrogen, USA) according to the manufacturer’s protocol. The real-time quantitative PCR mixture contained 2.0 μl of RT product, 10 μl of SYBR Green PCR mix (TransGen Biotech, China) and 0.5 mM primers in a final volume of 20 μl. The primers used in real-time PCR are listed S9 Table.