Fig. 5. Plk1 is mislocalized in NudC downregulated cells. HeLa cells were transfected with Luc siRNA or NudC siRNA for 72 hours. (A) Total cell lysates (10 µg) were immunoblotted with anti-NudC C peptide (top), anti-Plk1 (middle) or anti-ß-actin (bottom) antibodies. (B) Subcellular localization of Plk1. Cells were stained with anti-Plk1 antibodies and counterstained with DAPI as indicated. In Luc-siRNA-treated control cells, Plk1 staining was found on the centrosomes during prometaphase (a) and metaphase (c), and at the midbody during cytokinesis (e). By contrast, in NudC-siRNA-treated cells, Plk1 staining was diffuse and not localized to the centrosomes (g,i) or midbody (k). Notice the lagging DNA in the midzone during telophase in NudC-siRNA-treated cells (l, arrow). Stages in M phase were determined by DAPI stains of DNA (b,d,f,h,j,l). Bar, 10 µm for all panels.