Ctivity erh Ht and FAK cancer then remained high, indicating that the extract spindle checkpoint was sensitive. Despite the lack of mitotic spindles, ZM-treated extracts do not arrest in mitosis lie cdc2 activity t and chromosomes decondensed. In this particular experiment, the timing of the decline in H1 kinase activity t of histones in the extract ZM-contract in relation to the observed in the extract of contr On galvanized siege, But this deadline has not always seen. These results show that ZM either construction prevents the controlled station The spindle assembly, maintenance, or both. To investigate whether ZM affects establishment of the position of the contr Of CSF extract with 10,000 nuclei / l was erg Complements. As described above, the addition of calcium to induce the arrest, inactivation of cdc2 and chromosome decondensation.
When CSF extract was washed first with nocodazole and then incubated with calcium, remained histone H1 kinase activity FTY720 S1P Receptor inhibitor t high, and chromosomes decondensed, indicating that the extract was sensitive checkpoint. When CSF extract was added first with ZM and then incubated with nocodazole and calcium, lie Histone H1 kinase and chromosomes decondensed. In other experiments, cycling extracts in interphase first with ZM and then treated with nocodazole, were not arrested in mitosis. These results show that rt with ZM establishment of the controlled station st The integrity of t of the spindle. To test whether ZM affects maintenance of the controlled station Nocodazole was added at first, then ZM.
After the addition of calcium, remained histone H1 kinase activity t high, and the condensed chromosomes remained for the duration of the experiment. Similarly, when ZM was cycling extracts, which subsequently End of mitosis by nocodazole was added to arrest, was arrested extracts with high H1 kinase activity of t, and Figure 6 The addition of ZM to egg extracts does not prevent the formation of microtubules from sperm asters or centrosomes induced by Ran GTP, but reduce chromatin assembly induced by the spindle. ZM does not inhibit microtubule nucleation from centrosomes sperm. CSF extracts with sperm nuclei were erg Complements Min of rhodamine-labeled tubulin and then incubated on ice for 60 in the presence of either DMSO or ZM 20 to 21 M. The extracts were mentioned Rmt and samples were taken at intervals of 10 min and fixed.
Microtubules were visualized with rhodamine tubulin and DNA with Hoechst 33 042 by fluorescence microscopy. Enlarged TION, 60, 15 m. The figures show that the h Ufigsten structures occurring in extracts of DMSO and ZM from three independent Ngigen experiments were treated. ZM reduces spindle formation around chromatin-coated beads. Chromatin-coated beads were in CSF extract containing rhodamine-labeled tubulin and either DMSO or ZM 20 M. The samples were fixed at 60 min, and the status of the spindle formation was incubated determined. Enlarged TION, 60, 15 m. The images shown are the most hours Ufigsten structures occurring in extracts of DMSO pin and ZM-treated patients. The quantification of the structures around the spindle beads chromatin in DMSO and analysis of samples treated ZM example were fixed 60 min after initiation of spindle assembly. The nature of the structures with the spindle bead clusters that contain at least six beads were assigned to four associated