Quantitative real-time polymerase chain reaction (qPCR)

Total RNA was isolated from various samples and treated with RNAse-free DNase I. The DNase-treated RNA was reverse transcribed using random hexamer primers; the specific primers were designed to generate amplification fragments of 70–150 bp. Quantitative real-time polymerase chain reaction (qPCR) was performed on an ABI ViiA 7 Real-Time PCR platform using FastStart Universal SYBR Green Master with ROX in accordance with the manufacturer’s protocol; all reactions were performed in triplicate. The relative expression levels were calculated using the delta-delta Ct method, and HIS was used as a housekeeping gene [10].