*2.6. RT-qPCR Analysis*

The total RNA was extracted from cells using TRIzol Regent (Invitrogen, CA, USA) and purified by a treatment of DNase I (Thermo Fisher Scientific, Waltham, MA, USA). The high-quality RNA samples were used to generate the complimentary DNA (cDNA) using the PrimeScriptTM RT reagent Kit (TaKaRa, Japan). The PCR primers were designed by NCBI Primer BLAST (https://www.ncbi.nlm. nih.gov/tools/primer-blast/) and synthesized by Genscript Biotechnology (Nanjing, China) (Table S1). The Real-Time quantitative PCR (RT-qPCR) detection was performed using the SYBR Green master mix (Thermo Fisher Scientific, Waltham, MA, USA) according to manufacturer's instruction. Each PCR detection was performed with three biological and technical replicates. The gene expression values were normalized to an internal control (glyceraldehyde-3-phosphate dehydrogenase, GAPDH). The relative gene expression level was calculated as the relative quantification (RQ) using the 2–ΔΔCt method.
