Supporting Information

Files in this Data Supplement:

Fig. 5. Abnormal subcellular localization of endogenous PRC1 in Kif4 esiRNA-treated cells. HeLa cells transfected with control (luciferase) esiRNA or Kif4 esiRNA were fixed with 3% formaldehyde and then stained with rabbit anti-PRC1 antibodies (green), mouse anti-a -tubulin (red), and DAPI (DNA, blue). (Scale bar: 5 m m.)

Movie 1. Translocation of PRC1 during mitosis/cytokinesis. HeLa cells were transfected with EYFP-PRC1 and ECFP-H2B together with 100 nM control (luciferase) esiRNAs, and time-lapse images were collected at 2-min intervals for 10 h. The video was edited with SLIDEBOOK 3.0 software. EYFP-tubulin is shown in red (pseudocolored), and ECFP-H2B is shown in green (pseudocolored).

Movie 2. Abnormal translocation of PRC1 during mitosis/cytokinesis in cells treated with Kif4 esiRNA. HeLa cells were transfected with EYFP-PRC1 and ECFP-H2B together with 100 nM Kif4 esiRNAs, and time-lapse images were collected at 2-min intervals for 10 h. The video was edited with SLIDEBOOK 3.0 software. EYFP-tubulin is shown in red (pseudocolored), and ECFP-H2B is shown in green (pseudocolored).

Fig. 6. Localization of Kif4 and PRC1 in HeLa cells under the conditions of methanol fixation. HeLa cells grown on coverslips were fixed with methanol at -20°C for 5 min and stained with rabbit anti-Kif4 antibodies (green), Alexa Fluor 594-conjugated anti-PRC1 antibodies (red), mouse anti-a -tubulin antibody (blue), and DAPI (DNA, white). (Scale bar: 5 m m.)

Fig. 7. Colocalization of PRC1 and ectopically expressed GFP-Kif4 in Kif4 esiRNA-treated anaphase A cells. HeLa cells grown in a six-well plate were transfected with 400 ng of pEGFP-Kif4. Twenty-four hours later, the cells were transfected with 100 nM Kif4A esiRNA. Two days post-esiRNA transfection, cells were fixed and stained with anti-PRC1 antibodies (red) and DAPI (DNA, blue). The expression of GFP proteins is shown in green. A representative anaphase A cell is shown. (Scale bar: 5 m m.)