RNA extraction was conducted by employing TRIzol™ (Invitrogen), following the methods described before [27 (link)] with SYBR Green PCR Master Mix (Qiagen) for mRNA expression level determination and a Hairpin-it miRNAs qPCR kit (GenePharma) for miRNA expression level determination. The expression levels of GAPDH and RNU6B were used as endogenous controls. The primer sequences for PCR were shown in Table 1. Finally, the data were processed using the 2−ΔΔCt relative expression method. The more method details of real time PCR assay listed in Additional file 1.
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