Figure 3

TM and tail domains of NSP2 co-localize with aggresomes. A Confocal microscopy of Marc-145 cells mock infected or infected for 24 h with TA-12 and CH-1R at a multiplicity of infection of 0.01 (left margin), then stained for PRRSV (green) with anti-NSP2 antibodies and aggresomes (red) with aggresome detection reagent. The nucleus was stained with the DNA-binding dye DAPI (blue). Before harvesting, positive controls were set by incubation with diluted proteasome inhibitor (MG-132, 5 µM) for 18 h under normal cell culture conditions. B 293T and Vero E6 cells were transfected with NSP2-GFP, OTU-GFP, HV-GFP, TM-GFP, Tail-GFP, and empty vector (green). At 24 h later, cells were fixed and stained with aggresome detection reagent (red). The treatment method for positive cells is shown in (A). Yellow in the merged images represents the formation of aggresomes. Bar size: 25 µm.