FIGURE 8.

B23 functions in the recruitment of CENP-W to mitotic centromere during prophase. A, distribution of CENP-W and B23 in mitotic cells. After synchronization with nocodazole (100 ng/ml) for 12 h, HeLa-CENP-W cells were harvested at 30-min time intervals. Cells were then co-immunostained for CENP-W and B23 using anti-FLAG and anti-B23 antibodies, respectively. B, B23 depletion inhibits the centromeric recruitment of CENP-W. After incubation with B23 siRNA or control siRNA for 72 h, HeLa-CENP-W cells were incubated with nocodazole for 12 h. At 30 min after release, cells were harvested and CENP-W was immunostained. Anti-CENP-B, anti-CENP-C, and anti-centromere antibody (ACA, Cortex Biochem) were also used to visualize the centromere.