Figure 5

The teneurin-1 intracellular domain contains a nuclear localization signal. The amino acid sequence representing a putative nuclear localization signal (NLS) which was mutagenized to alanines (A). Western blot analysis of total lysates, cytoplasmic and nuclear fractions of COS-7 cells mock transfected, transfected with wild type ICD (ICD-WT) or transfected with the mutated NLS (NLS-Mut) constructs (B). Blots were probed with the anti-ICD, an antibody to the cytoplasmic protein vinculin, or an antibody to the nuclear protein Oct-1. Western blot quantification and calculation of the nuclear-to-cytoplasmic ICD ratio (C). Immunofluorescence staining of transfected COS-7 cells (D-G) and quantification of nuclear localization by cell counting (H). Both western blot analysis and immunofluorescence show that the overexpressed wild-type ICD is localized in the nucleus, whereas the NLS-Mut construct is predominantly found in the cytoplasm.