FIGURE 7:

LGN appears to be dispensable for anaphase cortical accumulation of NuMA. (A) Cortical intensity of endogenous LGN does not change from metaphase to anaphase. Representative images of metaphase (left) and anaphase (right) HeLa cells. Cells were fixed and stained with anti-LGN antibody (top) and DNA dye (bottom). Bar, 10 μm. (B) Quantification of polar cortical LGN fluorescence intensity from images in A. Results are from three independent experiments. Error bars, SD. n = 50 for each group of cells. (C, D) Knockdown of LGN did not prevent anaphase cortical accumulation of NuMA. MDCK cells stably expressing low level of Venus-NuMA were transfected with plasmids expressing control shRNA (top) or LGN-specific shRNA (bottom). At 72 h later, cells were either fixed and stained with anti-NuMA antibody (green) and DNA dye (blue; C) or subjected to time-lapse analysis (D) as described in Figure 5C. Bars, 10 μm.