Reverse transcription-qPCR (RT-qPCR)

Cells subjected to survivin shRNA lentivirus or control virus treatment for 24 h were lysed with TRIzol^® reagent (Ambion; Thermo Fisher Scientific, Inc.) and frozen at −80°C. RNA was extracted according to the protocol of the manufacturer. cDNA was synthesized using random six primers according to the protocol of the reverse transcription kit (cat. no. 240589; Tiangen Biotech, Beijing, China). qPCR was performed using a SYBR green-based protocol (Takara Bio, Inc., Otsu, Japan; cat. no. RR820L) (23). Primers were designed and synthesized by Sangon Biotech. (Shanghai, China). Relative gene expression was determined using the 2^−ΔΔCq method (23).