Figure 3.
Central spindle localized Aurora B is not essential for NuMA exclusion from the equatorial membrane. (A) Synchronization method for inactivation of Aurora B using ZM447439 to analyze cells in anaphase for NuMA and p150Glued localization, and after 12 h for assessing cytokinesis failure. (B and C) IF analysis of mitotically synchronized populations of HeLa cells as mentioned above either treated with DMSO (Control; B) or ZM447439 (C) for 12 h. Thereafter, cells were fixed and stained with anti-p150Glued antibodies (green). The scale bar in the panel and following panels represent 10 µm. Multinucleation percentage (%) is shown in the figure panel. (n > 500 cells were analyzed each from two independent experiments). (D and E) IF analysis of mitotically synchronized populations of HeLa cells that were either treated with DMSO (Control; D) or ZM447439 (E) for 30 min. Cells were fixed and costained with anti-NuMA (green) and anti-p150Glued (red) antibodies. More than 50 anaphase cells were analyzed from two independent experiments, and the representative cells are shown here. (F and G) IF analysis of HeLa cells in interphase that were either transfected with control siRNA (F) or siRNA-against Mklp2 (G). Cells were fixed after 40 h of siRNA transfection and stained with anti-p150Glued antibodies (green). Multinucleation percentage (%) is shown for each siRNA condition. (n > 500 cells from two independent experiments). (H and I) IF analysis of HeLa cells during anaphase that were either transfected with control siRNA (H) or siRNA-against Mklp2 (I). Cells were fixed after 40 h of siRNAs transfection and thereafter stained with anti-Aurora B antibodies (red). More than 30 anaphase cells were analyzed from two independent experiments, and the representative cells are shown here. (J and K) IF analysis of HeLa cells in anaphase that were either transfected with control siRNA (Control; J) or siRNA-against Mklp2 (K). Cells were fixed after 40 h of siRNA transfection and costained with anti-NuMA (green) and anti-p150Glued (red) antibodies. More than 50 anaphase cells were analyzed from two independent experiments, and the representative cells are shown here.

Central spindle localized Aurora B is not essential for NuMA exclusion from the equatorial membrane. (A) Synchronization method for inactivation of Aurora B using ZM447439 to analyze cells in anaphase for NuMA and p150Glued localization, and after 12 h for assessing cytokinesis failure. (B and C) IF analysis of mitotically synchronized populations of HeLa cells as mentioned above either treated with DMSO (Control; B) or ZM447439 (C) for 12 h. Thereafter, cells were fixed and stained with anti-p150Glued antibodies (green). The scale bar in the panel and following panels represent 10 µm. Multinucleation percentage (%) is shown in the figure panel. (n > 500 cells were analyzed each from two independent experiments). (D and E) IF analysis of mitotically synchronized populations of HeLa cells that were either treated with DMSO (Control; D) or ZM447439 (E) for 30 min. Cells were fixed and costained with anti-NuMA (green) and anti-p150Glued (red) antibodies. More than 50 anaphase cells were analyzed from two independent experiments, and the representative cells are shown here. (F and G) IF analysis of HeLa cells in interphase that were either transfected with control siRNA (F) or siRNA-against Mklp2 (G). Cells were fixed after 40 h of siRNA transfection and stained with anti-p150Glued antibodies (green). Multinucleation percentage (%) is shown for each siRNA condition. (n > 500 cells from two independent experiments). (H and I) IF analysis of HeLa cells during anaphase that were either transfected with control siRNA (H) or siRNA-against Mklp2 (I). Cells were fixed after 40 h of siRNAs transfection and thereafter stained with anti-Aurora B antibodies (red). More than 30 anaphase cells were analyzed from two independent experiments, and the representative cells are shown here. (J and K) IF analysis of HeLa cells in anaphase that were either transfected with control siRNA (Control; J) or siRNA-against Mklp2 (K). Cells were fixed after 40 h of siRNA transfection and costained with anti-NuMA (green) and anti-p150Glued (red) antibodies. More than 50 anaphase cells were analyzed from two independent experiments, and the representative cells are shown here.

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