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D box-dependent in vitro degradation of cyclin A, cyclin B, Xenopus geminin H, and S. pombe Cut2p is active in G1 and inactive in S phase. HeLa cells were synchronized in G1 or S phase. Cell extracts were prepared, supplemented with an ATP-regenerating system and 2.5 μM purified recombinant wild-type UbcH10, and then assayed for their destruction activity toward various in vitro translated proteins as indicated.
