Basic residues of the second AT-hook of HMGA2 are responsible for nuclear localization of fusion proteins

<p><b>Copyright information:</b></p><p>Taken from "The second AT-hook of the architectural transcription factor HMGA2 is determinant for nuclear localization and function"</p><p></p><p>Nucleic Acids Research 2007;35(6):1751-1760.</p><p>Published online 25 Feb 2007</p><p>PMCID:PMC1874589.</p><p>© 2007 The Author(s)</p> () Partial sequences of point-mutated constructs of HMGA2. Basic residues were replaced with alanine residues as indicated. Each point-mutant is fused C-terminally with EGFP (not drawn); numbers identify positions of residues on HMGA2 protein sequence. Summary of intracellular localization is indicated at right. () NIH-3T3 cells were transfected with HMGA2 point-mutant constructs, and fusion proteins were visualized by confocal laser microscopy. Propidium iodide (PI) staining of the same nuclei is shown. At least one hundred cells per transfection were analyzed in three different experiments, and at least one hundred cells per transfection were subjected to fluorescence quantification. Bars, 10 μm. () For each point-mutant construct, nuclear and cytoplasmic fluorescence were quantified, and percentage of nuclear localization was calculated. Bars indicate percentage ± standard deviation. () The expression and integrity of the fusion proteins were analyzed by Western blot using anti-GFP antibody