Fig. 5.

Pharmacological inhibition of p38 MAPK and JNK activation interferes with viral RNA synthesis. Vero cells pretreated with DMSO, SB202190, or SP600125 were mock-infected or infected with PEDV (MOI of 1) for 1 h and were incubated with either DMSO or each inhibitor. (A) Total cellular RNA was extracted at 48 hpi, and strand-specific viral genomic RNA (black bars) and sg mRNA (white bars) were amplified by quantitative real-time RT-PCR. Viral positive-sense genomic RNA and sg mRNA were normalized to mRNA of monkey GAPDH, and relative quantities (RQ) of mRNA accumulation were evaluated. The results on inhibitor-treated sample were compared with DMSO-treated results. The values shown are representative of the mean from three independent experiments, and the error bars denote standard deviations. *, P < 0.05; **, P < 0.001. (B) The extracted RNA was also subjected to northern blot analysis using the specific oligonucleotide biotin-labeled probes against the PEDV-specific 3′ UTR probe. Monkey GAPDH was served as an internal control to correct the data for variations in loading during viral RNA quantification. The positions of the genomic RNA and sg mRNAs are indicated next to the gel.