Subcellular distribution of DUX4 ΔNLS mutants.

<p>(<b>A</b>) DUX4 wild type (<i>WT</i>) as well as deletion mutants (<i>ΔNLS1</i>, <i>ΔNLS2</i>, <i>ΔNLS3</i>, <i>ΔNLS1-2</i>, <i>ΔNLS1-3</i>, <i>ΔNLS2-3</i> and <i>ΔNLS1-2-3</i>) were expressed in HepG2 cells and immunotsained using the monoclonal mAb9A12 antibody. Non background staining was observed when cells were transfected with the empty pcDNA3.1 vector (not shown; see Material and Methods section). (<b>B</b>) Western blot analysis of DUX4 wild type and NLS mutants showed in Fig. 2A, transiently expressed (i.e. 24 hs) in HepG2 cells. Cells transfected with an empty vector are shown (<i>vector</i>). The Western blot was developed using mAb9A12. The position of molecular weight markers (i.e. 72.8, 47.8 and 33.9 kDa) is indicated. (<b>C</b>) Percentage of nuclear-located DUX4 wild type and NLS mutants as determined by measuring the relative nuclear: cytoplasm fluorescence in HepG2 transfected cells (see Fig. 2A). Data are expressed as mean±SD of two independent experiments. The symbols (**) and (*) indicate significant difference <i>vs.</i> DUX4 wt, p<0.01 and p<0.05, respectively. For details, see text.