Fig. 7.

CsA impairs viral protein translation and viral RNA transcription. CsA-treated Vero cells were mock-infected or infected with PEDV (MOI of 0.1) for 1 h and further cultivated in the presence or absence of CsA. (A) At 48 hpi, cellular lysates were prepared, resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and immunoblotted using an antibody against the PEDV N protein. The blot was also reacted with anti-β-actin antibody to verify equal protein loading. PEDV N protein expression was quantitatively analyzed by densitometry in terms of the relative density value to the β-actin gene, and CsA-treated sample results were compared to DMSO-control results. (B) Total cellular RNA was extracted at 48 hpi, and strand-specific viral genomic RNAs (black bars) and sg mRNAs (white bars) of PEDV were amplified by quantitative real-time RT-PCR. Viral positive-sense genomic RNA and sg mRNA levels were normalized to monkey GAPDH mRNA, and relative quantities (RQ) of mRNA accumulation were evaluated. CsA-treated sample results were compared with DMSO-treated results. Values are representative of the mean from three independent experiments and error bars denote standard deviations. *, P=0.001 to 0.05; †, P<0.001.