Quantitative real-time PCR (qPCR)
Total RNA of the PEDV-infected cells or recombinant plasmid-transfected
cells was extracted using RNAiso Plus reagent (Takara, Japan) according
to the manufacturer’s protocols. The concentrations of the extracted RNA
were measured using a Thermo Scientific Nano Drop 2000c system (Thermo
Scientific, USA). Gene-specific primers for qPCR are listed in Table S3.
qPCR was performed using ChamQ Universal SYBR qPCR Master Mix on an ABI
7300 real-time PCR system. β -actin served as the endogenous
control. The relative values for mRNA accumulation were determined using
the 2-ΔΔCt method and compared with mock-treated
results22. We tested all compounds independently five
times to obtain five samples (N=5), and each sample was run in
triplicate.