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. 2005 Mar 7;102(12):4318–4323. doi: 10.1073/pnas.0501108102

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Copyright © 2005, The National Academy of Sciences

Freely available online through the PNAS open access option.

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Fig. 4. — Emi2 is modified, apparently ubiquitinated by SCF^βTrCP, and destroyed in CSF extract after calcium addition. (A) Emi2 is destroyed during CSF release. Full-length, radiolabeled IVT Emi2 was incubated in CSF extract with or without Ca²⁺ addition or in interphase extract for the indicated times. (B) Emi2 is destroyed through its conserved βTrCP recognition degron. Emi2 or mutants in one (DS33AA or DS283AA) or both (2xDS-AA) candidate degron sites were incubated in CSF extract and destruction was assayed at the indicated times after calcium addition. (C) Addition of calcium triggers Emi2 binding toβTrCP in CSF extract. IVT myc-Emi2 and radiolabeled IVT βTrCP were incubated in CSF extract with proteasome inhibitors, with or without calcium addition, for the indicated times. Anti-myc immunoprecipitates were analyzed for bound βTrCP.