Expression of VZV ORF25. (A) Immunoblot analysis was performed with the ORF25 rabbit polyclonal antiserum on mock infected or VZV infected IMR90 cell extracts. (B) Indirect immunofluorescence analysis of pORF25 expressed in transiently transfected cells. Cells were transfected with pcDNA3.1D/V5-ORF25 and 48 hr post-transfection fixed, stained with DAPI and incubated with an anti-V5 primary antibody followed by a FITC conjugated secondary antibody. (C) Indirect immunofluorescence analysis of pORF25 expressed in HSV-2 infected cells. Cells were transfected with pcDNA3.1D/V5-ORF25 and infected with HSV-2 24 hr post-transfection. Transfected, infected cells were fixed at 10 hr post-infection, stained with DAPI, and incubated with an anti-V5 rabbit and anti-HSV-2 ICP8 mouse antibodies followed by FITC anti-rabbit and rhodamine red-X conjugated anti-mouse secondary antibodies. pORF25 expressing cells appear green, HSV-2 infected cell nuclei appear red and DAPI stained nuclei appear blue.