FIG. 6.

Suppression of in vivo p53 acetylation by vIRF expression. (A) Western blot assay of in vivo p53 acetylation. Saos-2 cells were infected with recombinant Ad-p53 and/or Ad-vIRF as indicated at the top. At 48 h postinfection, cell lysates were used for immunoprecipitation (IP) with anti-p53, anti-p53(Ac320), and anti-p53(Ac373) antibodies, followed by Western blot (WB) analysis with the horseradish peroxidase-conjugated anti-pan-p53 antibody. The data were reproduced in two independent experiments. (B) Immunofluorescence test of in vivo p53 acetylation. The Saos-2 cells described above were stained with anti-p53(Ac320) and anti-p53(Ac373) antibodies. Cells were visualized with Nomarski optics. Equivalent levels of p53 were detected in both cells with an anti-p53 antibody (see above). The immunofluorescence test was performed with a Leica confocal immunofluorescence microscope.