RNA Isolation and qPCR Analysis

RNA was isolated using the RNeasy or miRNeasy Kit (Qiagen, Hilden, Germany) according to the manufacturer’s recommendations. The concentration of total RNA was determined by NanoDrop ND-1000.
cDNA for mRNA analysis was prepared using the RevertAidTM H minus First-strand Kit (Thermo Fischer Scientific, Massachusetts, USA). Primers and probes are listed in Additional file 2: Supplementary Table 3. For quantification of miRNAs, miScript RT and PCR system (Qiagen, Hilden, Germany) was used. Raw data analysis was performed by using QuantStudio PCR Systems (Applied Biosystems). Data acquisition and raw data analysis were performed using QuantStudio PCR Systems (Applied Biosystems) with the ΔΔCt method [24 (link)].