Supplemental Materials

This article contains the following supporting material:

• Supplemental figure 1. - Kif12 null cells have a grossly normal central spindle and normal chromosome segregation during anaphase. (A, B) The central spindle of Kif12 null is grossly normal. Immunofluorescence images of Kif12 null cells stained with DAPI and anti-tubulin antibody are shown. DNA is shown in blue and microtubules are shown in red. (C) Time lapse fluorescence images of a Kif12 null cell expressing GFP-histone H2B during mitosis show that chromosome segregation is normal in anaphase. No lagging chromosomes or other aberrations were observed in Kif12 null cells. Times are indicated in minutes: seconds. Bars, 5 µm.
• Supplemental figure 2. - Kif12 is essential for the localization of DdINCENP_1-500 at the cleavage furrow. GFP-DdINCENP _1-500 was expressed in Kif12 null and wild type cells separately. Time lapse fluorescence micrographs of these cells during cytokinesis are shown. Arrow heads point to the cleavage furrow. GFP- DdINCENP_1-500 localized to the cortex of the cleavage furrow in wild type cells (C) (See Movie 9). However, it was not found at the cleavage furrow in Kif12 null cell (A, B) (See Movie 10). Times are indicated in minutes: seconds. Bars, 5 µm.
• Movie01 - Kif12 localizes to the cleavage furrow during cytokinesis. A time lapse fluorescence microscopy video of a kif12 null cell expressing GFP-Kif12 during cytokinesis. GFP-Kif12 localizes to the cortex of the cleavage furrow in additional to the central spindle. Moreover, it highly concentrates to the cytoplasmic bridge connecting the daughter cells during abscission. Frames were acquired every 20 s and are shown at 3 frames/s. Time is indicated in minute: second. Also see Figure 1.
• Movie02 - Time lapse fluorescence microscopy video of a wild type cell expressing GFP-DdINCENP during cytokinesis. DdINCENP began to localize to the cortex region of the cleavage furrow at the beginning of the cytokinesis and highly concentrated on the cytoplasmic bridge during the abscission. This pattern of localization is similar to that of Kif12. Frames were acquired every 15 s and are shown at 3 frames/s. Time is indicated in minute: second. Also see Figure 2 (A, B).
• Movie03 - Time lapse fluorescence microscopy video of a kif12 null cell expressing GFP-DdINCENP during cytokinesis. Although GFP-DdINCENP still localizes to the central spindle and spindle pole bodies, it was no longer found at the cleavage furrow in the absence of Kif12 (compared to Movie 2). Furthermore, GFP-DdINCENP was also absent from the cytoplasmic bridge during the abscission, while it highly concentrated on the bridge in wild type cells (see Movie 2). Time is indicated in minute: second. Frames were acquired every 15 s and are shown at 2 frames/s. Also see Figure 2.
• Movie04 - Time lapse fluorescence microscopy video of a DdINCENP null cell expressing GFP-Kif12 during cytokinesis. In DdINCENP null cells, GFP-Kif12 was still found at the cortex of the cleavage furrow from the early stage of cytokinesis and continued to build up at the furrow area. At the end of the cytokinesis, GFP-Kif12 was highly enriched at the cytoplasmic bridge (also see Figure 3). Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s.
• Movie05 - Time lapse fluorescence microscopy video of a DdINCENP null cell expressing GFP- DdINCENP_488-1320 during cytokinesis. DdINCENP_488-1320, was absent from the cortex of the cleavage furrow and was homogenously distributed in the cytoplasm during cytokinesis. Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s. Also see Figure 5.
• Movie06 - Time lapse fluorescence microscopy video of a DdINCENP null cell expressing GFP- DdINCENP_1-1013 during cytokinesis. DdINCENP_1-1013 localizes to the central spindle and the cleavage furrow despite the lack of an IN-box sequence. Furthermore, it also concentrates to the cytoplasmic bridge during abscission. Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s. Also see Figure 5.
• Movie07 - Time lapse fluorescence microscopy video of a DdINCENP null cell expressing GFP- DdINCENP_1-500 during cytokinesis. DdINCENP_1-500 also localizes to the cleavage furrow. Noticeably, it highly concentrates to the cytoplasmic bridge during abscission. Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s. Also see Figure 5.
• Movie08 - Time lapse fluorescence microscopy video of a DdINCENP null cell expressing GFP- DdINCENP_1-273 during cytokinesis. DdINCENP_1-273 still localizes to the cleavage furrow, although it has a more diffuse distribution in the cytoplasm than DdINCENP_1-1013 does. Nevertheless, it still highly concentrates to the cytoplasmic bridge. Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s. Also see Figure 5.
• Movie09 - Time lapse fluorescence microscopy video of a wild type cell expressing GFP- DdINCENP_1-500 during cytokinesis. DdINCENP_1-500 localizes to the cleavage furrow in wild type cells. Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s.
• Movie10 - Time lapse fluorescence microscopy video of a Kif12 null cell expressing GFP- DdINCENP_1-500 during cytokinesis. Similar to the full length DdINCENP, DdINCENP_1-500 depends on Kif12 to localize to the cleavage furrow (compared to Movie 9). In the absence of Kif12, DdINCENP_1-500 didn't localize to the cleavage furrow. Nor was it found at the cytoplasmic bridge. Times are indicated in minutes: seconds. Frames were acquired every 20 s and are shown at 3 frames/s.
• Movie11 - Two time lapse fluorescence microscopy videos of wild type cells expressing GFP- DdINCENP_1-500 during interphase. GFP- DdINCENP_1-500 localizes to the cortex of the plasma membrane during interphase. Furthermore, it was enriched at the bottom of pinocytic cups. Times are indicated in minutes: seconds. Frames were acquired every 25 s and are shown at 3 frames/s.
• Movie12