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Fig. 6. Destruction of purified, recombinant human Aur-A protein in Xenopus egg extract requires the D box and sequences within the A box. Destruction assays were performed in Xenopus egg extracts as follows. Xenopus egg CSF extract was incubated for 5 minutes with the APC/C activators Cdc20 or Cdh1. His6-tagged human Aur-A wild-type (A) or mutant (B) protein substrates were added at a final concentration of 220 nM and incubated for a further 10 minutes. Calcium was added to trigger exit from meiosis-II metaphase arrest and entry into the first cell cycle. Samples were taken at the indicated times and analysed SDS-PAGE and western blotted with anti-Aur-A antibodies.





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