FIG 5.
PDCoV nsp5-induced cleavage of pDCP1A impairs its antiviral activity. (A) Expression of wild-type pDCP1A and pDCP1A mutants in IPI-2I cells. Wild-type pDCP1A, pDCP1A-Q343A, pDCP1A1–343, or pDCP1A344–580 was transfected into IPI-2I cells and analyzed by Western blotting after 24 h. (B) IPI-2I cells were cotransfected with PDCoV nsp5 along with wild-type pDCP1A or pDCP1A-Q343A and collected after 28 h for Western blotting. (C) Flag-tagged pDCP1A or pDCP1A-Q343A mutant expression plasmids were transfected into IPI-2I cells. After 12 h, cells were infected with PDCoV (MOI = 0.5) for 24 h. The cells were harvested and lysed for Western blot analysis. (D and E) IPI-2I cells were transfected with expression plasmid encoding wild-type pDCP1A, pDCP1A-Q343A, pDCP1A1–343, or pDCP1A344–580. After 24 h, cells were infected with PDCoV (MOI = 0.5). PDCoV genomic RNA and total RNA or viral titer was detected by RT-qPCR (D) or TCID50 assay (E), respectively, at 24 hpi. ns, not significant; *, P < 0.01.