Figure 3

A. Localization of BVDV NS4B in infected MDBK cells. Cells were grown on coverslips and infected with BVDV at MOI of 10. At 12 h (i and ii), 18 h (iii and iv) and 24 h.p.i (v and vi), cells were processed for immunofluorescence (IF) with NS4B-specific antibody (1/50 dilution) and Alexa fluor 488-conjugated secondary antibody (1/500). Nuclei were stained with DAPI. Notice the Golgi-like NS4B distribution at 12 h and 18 h.p.i., whereas foci are seen at 24 h.p.i No fluorescence is displayed in mock-infected cells (vii). Bars = 10 μm. B. BVDV NS4B partially colocalizes with Golgi markers. Cells were grown on coverslips and infected with BVDV as above. At 18 h.p.i., cells were processed for IF with NS4B- (ii and iv; vi and viii), TGN38- (iii and iv) and Golgin97 (vii and viii)- specific antibodies. Notice the colocalization of NS4B with TGN38 or Golgin97 protein. Mock-infected cells, stained with anti TGN38 (i) or Golgin97 (v), are also shown.