RNA Extraction and qRT-PCR Analysis

Total RNA was extracted using the Pure Link^TM RNA Mini Kit (Invitrogen, California, CA, USA), according to the manufacturer’s instructions [8 (link),9 (link)]. The quantification of total RNA was performed by a Nano-MD UV-Vis spectrophotometer (Scinco, Seoul, Korea). Complementary DNA (cDNA) was synthesized using RevertAid Reverse transcriptase (Thermo Scientific, Waltham, MA, USA) in a 20 μL reaction volume. Quantitative reverse transcription (qRT) polymerase chain reaction (PCR) was performed and analyzed using TB Green™ Premix Ex Taq™ (Takara, Shiga, Japan) and Thermal Cycle Dice real-time PCR system (Takara, Shiga, Japan) as reported previously [9 (link)]. Three independent replicates were performed for each primer sets. The primer sequences used in this study are provided in Table S1.