ORF6 interacts with KPNB1. (A) ORF6 colocalization with KPNB1. Vero cells were transfected with Flag-tagged KPNB1 and HA-ORF6. At 24 h posttransfection, cells were fixed and stained. Cells were incubated with mouse anti-HA (αHA) and rabbit anti-Flag antibodies. Anti-Flag antibody was visualized with Alexa Fluor 546-conjugated secondary antibody, and anti-HA antibody was visualized with Alexa Fluor 488-conjugated secondary antibody. Coverslips were visualized with a Zeiss confocal microscope. (B) 293 cells were transfected with HA-tagged ORF6 and Flag-tagged KPNA1, KPNA2, KPNA3, or KPNA4. Cells were lysed 24 h posttransfection and immunoprecipitated (IP) with anti-HA antibody. Proteins were separated on a 4 to 12% SDS PAGE gel and analyzed by Western blot (WB) probing with anti-HA, anti-Flag, and anti-KPNB1 antibody.