FIG. 6.

Nutlin-3a disrupts LANA:hdm2:p53 complexes in vivo and in vitro. (A) BC-3 cells were treated with nutlin-3. p53, hdm2, and LANA levels and complexes were determined by Western blotting (input) and immunoprecipitation followed by Western blotting. Also shown are results of Western blotting for p21 levels to document p53 and caspase-3 activation, with actin as a loading control. (B) The disruption of LANA:p53:hdm2 interaction by nutlin-3 is a direct effect. Purified LANA:p53:hdm2 complexes from heparin fractions 3 to 7 were pooled and incubated with increasing amounts of nutlin-3 in vitro; p53 complexes were immunoprecipitated (IP) and blotted (WB) with LANA or p53 antibodies.