Total RNA was isolated using TRIzol®Plus RNA Purification Kit (Invitrogen) and RNase-Free DNase Set (Qiagen) followed by reverse transcription using the SuperScript™ III First-Strand Synthesis SuperMix (Invitrogen) strictly according to the manufacturer’s instructions. Real-time qPCR was performed in triplicate on an ABI Prism 7700 Sequence Detector system (Applied Biosystems, Foster City, CA, USA) using an annealing temperature of 63°C and gene-specific primers listed in Table 1 . The data were normalized to GAPDH or 18S rRNA and calculated by 2−ΔΔCT method (27 (link)).
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