Extraction and Quantification of Total RNA

Total RNAs were isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) as previously described in our recent study [32 (link)]. A NanoDrop One^C spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) was used to ascertain RNA concentration. First-strand cDNA was synthesized from 1.0 μg of total RNA in 20 μL volume using the PrimeScript reverse transcriptase (RT) kit (containing gDNA Eraser, Perfect Real Time, TaKaRa, Dalian, China) according the manufacturer’s recommendations. The cDNA was diluted 10-fold for the subsequent general polymerase chain reaction (PCR), reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and dsRNA synthesis experiments.