Fig. 4
From: Adducin family proteins possess different nuclear export potentials
The leucine-rich motif in the neck domain is a functional NES for ADD1, but not for ADD2 and ADD3. a A leucine-rich motif (FEALMRMLDWLGYRT) is conserved in all three ADD isoforms. The tail domain alone and the fragment containing both neck and tail domains (neck + tail) with or without the deletion of the leucine-rich motif were constructed. These constructs were transiently expressed in HEK293 cells and their expression was analyzed by immunoblotting with anti-HA antibody. b The constructs described in panel a were transiently expressed in NIH3T3 cells and their distribution in the nucleus and cytoplasm was measured (n > 300). The values (mean ± SD) are from three experiments. c The full-length HA-ADD proteins and their respective mutants with the deletion of the leucine-rich motif (FEALMRMLDWLGYRT) in the neck domain were transiently expressed in HEK293 cells and their expression was analyzed by immunoblotting with anti-HA antibody. d The constructs described in panel c were transiently expressed in NIH3T3 cells and their distribution in the nucleus and cytoplasm was measured (n > 600). The values (mean ± SD) are from three experiments. e The Φ-X3-Φ-X 2-Φ-X1-Φ motif ("Φ" is a hydrophobic residue) was found in the neck domain of ADD2 and ADD3. HA-ADD2, HA-ADD3, and their respective mutants with the deletion of this motif were transiently expressed in HEK293 cells and their expression was analyzed by immunoblotting with anti-HA antibody. f The constructs described in panel e were transiently expressed in NIH3T3 cells and their distribution in the nucleus and cytoplasm was measured (n > 600). The values (mean ± SD) are from three experiments