FIG. 2.

Colocalization of vIRF with p53. (A) Colocalization of vIRF with p53 in KSHV-infected BCBL-1 cells. KSHV-infected BCBL-1 cells were fixed and reacted with rabbit polyclonal anti-vIRF and mouse monoclonal anti-p53 antibodies. vIRF protein was detected with an anti-rabbit secondary antibody conjugated with Alexa 568 (red), and p53 protein was detected with an anti-mouse secondary antibody conjugated with Alexa 488 (green). Cells were visualized with Nomarski optics after Topro-I nuclear staining (blue). These two panels are representatives of 10 different fields. (B) Colocalization of vIRF with p53 in COS-1 cells. COS-1 cells were transfected with expression vector containing the Flag-tagged vIRF gene. At 48 h posttransfection, cells were fixed and reacted with mouse monoclonal anti-Flag and rabbit polyclonal anti-p53 antibodies. p53 protein was detected with an anti-rabbit secondary antibody conjugated with Alexa 488 (green), and the Flag-tagged vIRF protein was detected with an anti-mouse secondary antibody conjugated with Alexa 568 (red). Cells were visualized with Nomarski optics after Topro-I nuclear staining (blue). The immunofluorescence test was performed with a Leica confocal immunofluorescence microscope. The yellow color in merged panels indicates colocalization of the red and green labels. The data were reproduced in three independent experiments.