Figure 3.
Figure 3. Artificial retargeting of Aurora B restores MCAK and CENP-AS7ph at centromeres of Haspin-inhibited cells. (A and B) HeLa cells were transfected with CENP-B–EGFP or EGFP–CENP-B–INCENP plasmids between double thymidine treatments. 7 h after release from G1/S, 30 nM nocodazole was added to accumulate mitotic cells. Then, 3.5 h later, medium containing 20 µM MG132 with or without 10 µM Haspin inhibitors or 50 nM Hesperadin, in the presence (A) or absence (B) of 0.33 µM nocodazole, was added for 75 min. Approximately 100 mitotic cells in each condition from one experiment were classified according to the intensity of centromeric MCAK (A) or CENP-AS7ph (B) by immunofluorescence microscopy. Similar results were obtained in a second experiment. Bars, 5 µm.

Artificial retargeting of Aurora B restores MCAK and CENP-AS7ph at centromeres of Haspin-inhibited cells. (A and B) HeLa cells were transfected with CENP-B–EGFP or EGFP–CENP-B–INCENP plasmids between double thymidine treatments. 7 h after release from G1/S, 30 nM nocodazole was added to accumulate mitotic cells. Then, 3.5 h later, medium containing 20 µM MG132 with or without 10 µM Haspin inhibitors or 50 nM Hesperadin, in the presence (A) or absence (B) of 0.33 µM nocodazole, was added for 75 min. Approximately 100 mitotic cells in each condition from one experiment were classified according to the intensity of centromeric MCAK (A) or CENP-AS7ph (B) by immunofluorescence microscopy. Similar results were obtained in a second experiment. Bars, 5 µm.

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