Fig. 2.

Determination of PEDV ORF3 protein expression characteristic. (A) HeLa cells were transiently transfected with indicated plasmids for 24 h, respectively. Cell lysates were harvested and subjected to Western Blot probed with either mouse monoclonal anti-Flag (left panel) or anti-Myc (right panel) antibodies followed by the goat-anti-mouse-HRP antibody. β-actin was detected to serve as internal loading control. (B) HeLa cells were transiently transfected with indicated plasmids for 24 h, respectively. Cells were fixed, permeabilized and then probed with mouse monoclonal anti-Flag (upper panel) or anti-Myc (lower panel) antibodies, followed by FITC-labeled goat-anti-mouse (green) or TRITC-labeled goat-anti-mouse (red) antibodies with confocal microscope analysis. Nuclei were identified by DAPI staining. (C) Three types of cells, including Vero, 293 T and PK15, were either transfected with pEGFP-N1 or pEGFP-N1-ORF3 for 24 h, respectively. Cells were fixed, permeabilized, and directly examined using a GFP filter of confocal immunofluorescence. Nuclei were identified by DAPI staining. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).