2.8. Quantitative Real-Time PCR

Total RNA was extracted by TRIzol reagent (Invitrogen, USA). According to the reverse transcription kit (TaKaRa, Japan) instructions, reverse transcription reaction was performed to synthesize cDNA strands, and SYBR green (TaKaRa, Japan) was added for qRT-PCR amplification on ABI 7500 System (ABI, USA). U6 or GAPDH was used as internal reference, and the relative expression of RNAs was calculated by the 2^−ΔΔCt method. Primers are listed in Table 1.

Primer sequences for real-time fluorescence quantification PCR.