Table I.
Construct | Nucleus (percentage of cells) | Cytoplasm (percentage of cells) | Number of cells | |||
---|---|---|---|---|---|---|
β-gal | 100 | 0 | 49 | |||
β-gal-Zyx305-411 | 5 | 95 | 98 | |||
β-gal-Zyx305-351 | 20 | 80 | 85 |
To test the ability of zyxin's conserved leucine-rich region to relocalize nuclear proteins to the cytoplasm, the distribution of β-gal with and without fused zyxin amino acid sequences was determined by immunofluorescence. Plasmids engineered to express β-gal or β-gal–zyxin fusion proteins were injected into REF-52 cells, and cells were stained with anti–β-gal antibodies 24 h after injected. Cells that expressed the construct were identified by fluorescence microscopy, and the distribution of fusion protein was scored. Results from three experiments are shown.