FIG 6.

Effect of Rabs on PDCoV infection. (A) Effect of DN Rab proteins on PDCoV infection was determined via confocal microscopy. Cells transfected with plasmid constructs encoding Flag-tagged Rab5, Rab7, Rab11 WT, or DN were infected with PDCoV (MOI of 5) at 37°C for 6 h. Cells were fixed with 4% paraformaldehyde and then reacted with anti-Flag and anti-PDCoV N antibodies and visualized via confocal microscopy. Bar, 10 μm. (B to D) Rab5 and Rab7, but not Rab11, knockdown inhibited PDCoV infection. siRab5- (B), siRab7- (C), siRab11- (D), or siCtrl-transfected cells were infected with PDCoV (MOI of 5) for 6 h. Cells were lysed to determine the silencing efficiency of Rabs, viral RNA copy numbers, and N protein expression levels via RT-qPCR and Western blot analysis. Target protein expression was quantitatively estimated by ImageJ software and presented as the density value relative to that of the β-actin. The presented results represent the means and standard deviations of data from three independent experiments. *, P < 0.05; **, P < 0.01; ns, nonsignificant difference.