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. 2005 Jul 29;6(9):885–890. doi: 10.1038/sj.embor.7400487

Figure 3.

Figure 3

Nuclear retargeting of an M6 mutant restores its intercellular transfer. (A–C) HeLa cells transfected with KNHD (A), KNM6HD (B) or KNM6HD:NLS (nuclear localization signal; C) were cultured for 24 h, fixed and immunostained with an anti-haemagglutinin (anti-HA) antibody. Compared with KNHD (A) and KNM6HD:NLS (C), which mainly localize in the nucleus, KNM6HD accumulates in the cytoplasm (B). (D) HeLa cells transfected with KNM6HD:NLS were processed as in Fig 1A. Addition of an ectopic NLS to KNM6HD restores intercellular transfer. (E) Membrane fractions from cells transfected with the indicated constructs were separated on an Optiprep (10%–30%) density gradient and analysed by western blot with an anti-HA antibody. Although expressed at similar levels, KNM6HD is weakly detected in the different fractions compared with KNHD and KNM6HD:NLS (1, low density; 10, high density). The arrows represent transfected HeLa cells. Scale bars, 50 μm.