Fig. 2.

The AnkG antiapoptotic domain mediates binding to the proapoptotic protein p32. (A) CHO cells ectopically expressing the indicated HA-tagged truncations of AnkG were incubated with staurosporine. The cells were stained with DAPI and HA-specific antibody. The nuclear morphology of HA-expressing cells was scored as in Fig.1A. *P < 0.003. (B) HEK 293 cells were cotransfected with plasmids encoding GFP-tagged p32 and the indicated HA-tagged truncations of AnkG. Proteins were precipitated from the cell lysates with an anti-GFP antibody. Immunoblot analysis was used to detect p32 (α-GFP) and AnkG (α-HA) in the lysates and immunoprecipitates (IP). (C) HeLa cells transfected with control siRNA (control), with p32 siRNA (p32), or untransfected (mock) were exposed to UV light as indicated. The cells were stained with DAPI, and nuclear morphology was scored. The percentage of cells with apoptotic nuclear morphology was calculated. One hundred nuclei per sample from three independent experiments were counted. *P < 0.006.