Total rna isolation reagent

For real-time quantitative RT-PCR (qPCR) analyses, total RNA from the indicated cells was extracted using the total RNA isolation reagent (Promega, Madison, WI, USA). Reverse transcription was performed with the PrimeScript RT reagent kit (TaKaRa, Dalian, Liaoning, China) using 1 ìg of total RNA. SYBR Green qPCR Master Mixes (ThermoFisher Scientific, Waltham, MA, USA) were used for qPCR detection of the cDNA, and qPCR reactions were performed on ViiA^TM 7 Dx system (Applied Biosystems, Foster, CA, USA). The level of target genes was normalized to the expression of an internal control gene (rpoD), which yielded a 2^−ΔΔCt value. Sequences for primers are listed in Appendix 1.

Total RNA was extracted using total RNA isolation reagent (Promega, Madison, WI, USA). Reverse transcription (1 μg of total RNA) was performed with the PrimeScript RT reagent Kit (Takara, Dalian, Liaoning, China). The cDNA was subjected to qPCR on a ViiA™ 7 Dx system (Applied Biosystems, Foster, CA, USA) using SYBR Green qPCR Master Mixes (ThermoFisher Scientific). The expression levels of the target genes were normalized to the expression of the internal control gene (rpoD), using the 2^−ΔΔCt method. The sequences of the primers are listed in Table S1.