binding to Mad2 protein. (a) Diagrams of N-terminal deletion p31comet fragments. (b) p31comet binding to Mad2 protein. EGFP, EGFP-tagged full-length p31comet, and a series of N-terminal deletion EGFP-tagged p31comet were overexpressed in HeLa cells by adenovirus infection and the expression level of each protein was monitored by western blotting with anti-GFP antibody, using anti-Mad2 and anti-EB1 antibodies for loading controls. The cell extracts overexpressing each protein were prepared for immunoprecipitation with anti-GFP antibody, followed by western blotting with anti-Mad2 and anti-GFP antibodies. (c) Fluorescence-activated cell sorting (FACS) analysis of the cells overexpressing EGFP-tagged full-length p31comet and a series of N-terminal deletion EGFP-tagged p31comet in the presence of nocodazole (100 ng/mL). A population of >4 N DNA content in the cells was shown.