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. 2005 Nov 1;102(46):16853–16858. doi: 10.1073/pnas.0507365102

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Copyright © 2005, The National Academy of Sciences

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Fig. 2. — PH domain of PIKE dictates its nuclear localization. (A) PIKE-L PH domain resides in the nucleus. GFP-tagged PH, PZA, and full-length PIKE-L were transfected into HEK293 cells. WT proteins and dominant-negative PIKE-L (K413A,S414N) predominantly distribute in the nucleus. The nuclei were stained with DAPI (Lower, leftmost image). By contrast, PH domain and PZA fragment mutants, unable to bind phosphatidylinositols, exclusively localize in the cytoplasm, whereas PIKE-L (K679,687N) distributes in both the cytoplasm and the nucleus. GST-tagged PH domain also exclusively resides in the nucleus (Upper, rightmost image). (B) PIKE-A PH domain and its full-length proteins localize in the cytoplasm, no matter whether it binds phosphatidylinositol lipids or not (leftmost images). (C) The expression of GST- and GFP-tagged constructs is verified.