PEDV N expression enhances reverse-genetics rescue from pPEDV-mCh.

HEK 293T cells were co-transfected with pPEDV-mCh (RG PEDV-WT) or pPEDV-mChN.L381P (RG PEDV-mt) and pCAGGS-N (or an empty vector). At 48 hpt, cells were lysed and analyzed by Western blotting with anti-PEDV N antibody (A) and supernatants were harvested and adsorbed onto Vero E6 cells. At 48 hpi, fluorescence images were taken to monitor replication of reverse genetics-derived PEDV-mCh (B) or PEDV-mChN.L381P (C) in Vero E6 cells. Supernatants were harvested for viral titer determination by RT-qPCR with M-specific primers (B and C). Experiments were repeated with pCAGGS expressing PEDV N derived from field strains (D). Values are averages±SEM of three independent experiments. GE, genome equivalents. * p p < 0.01.