Figure 8. Kinetochore Borealin can be uncoupled from pH2A^T120.

HeLa M cells were synchronized and treated as shown in the schematic. Briefly, cells were transfected with the indicated Borealin-FKBP and synchronously released into a taxol block. All cells received AP20187 to induce dimerization. Once cells were in mitosis, 5Itu was added for 90 minutes. Then, either ZM447439 (ZM) or reversine (Rev) was added for an additional 30 minutes before cells were fixed. Immunofluorescence was carried out with antibodies to pH2A^T120, Hec1, or HA to detect the Borealin-FKBP fusion. Maximum projection images are shown (scale bar = 10μm).