Fig. 2. Deletion mutagenesis mapping of the p120^ctn NLS. (A) p120^ctn deletion constructs for NLS analysis. Each p120^ctn fragment analyzed was fused N-terminally to ß-gal and C-terminally to GFP. (B) Using HeLa cells, nuclear localization of our p120^ctn deletion mutants was scored relative to the ß-gal-GFP fusion negative control and the SV40-LTAg-NLS-ß-gal-GFP positive control. (C) Forced expression of full-length p120^ctn fused to ß-gal and GFP resulted in a nuclear and cytoplasmic localization of the fusion protein. (D) Of our five p120^ctn deletion mutants, one mutant (p120^ctn-2) showed nuclear localization in HeLa cells. However, this deletion mutant also localized to distinct cytosolic aggregates. (E) A putative NLS, encoded by amino acids 306-320 of p120^ctn, does not show NLS activity as it was insufficient to target a ß-gal-GFP fusion to HeLa nuclei. The images shown are representative of at least 100 cells observed for each construct. Bars, 10 µm.