FIG 6.

Mutation of Thr56 and Thr61 impairs nuclear localization of apoptin in H1299 cells. (A) H1299 cells were transfected with Flag-tagged WT or mutant apoptin. The cells were allowed to proliferate for 24 h prior to processing for Flag immunofluorescence with nuclear staining (DAPI). (B) Quantification of cells displaying nuclear apoptin was determined by scoring at least 100 Flag-positive cells per condition. **, P ≤ 0.01. The error bars indicate SEM. (C and D) H1299 cells were transfected with the indicated constructs for 24 h. Whole-cell extracts were prepared for immunoprecipitation of Flag complexes and immunoblot analysis with the indicated antibodies. E.V. indicates empty vector.