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. Author manuscript; available in PMC: 2011 Oct 19.
Published in final edited form as: Cancer Cell. 2010 Oct 19;18(4):382–395. doi: 10.1016/j.ccr.2010.08.010

Figure 1. TAME inhibits APC activation by perturbing binding of Cdc20 or Cdh1.

Figure 1

(A) Structures of TAME and AAME. (B) TAME induces mitotic arrest in Xenopus extract. Recombinant cyclin B1/Cdk1 was added to interphase extract in the presence of compounds. Cdc27 phosphorylation and cyclin B1 levels were examined by immunoblot. (C) TAME inhibits APC activation. Compounds were added to mitotic Xenopus extract immediately before APC immunoprecipitation. The activity of the isolated APC was measured in a reconstituted assay. (D) TAME inhibits Cdc20 association with mitotic APC. Compounds were added to mitotic Xenopus extract prior to APC immunoprecipitation. Numbers represent CCD-imaging based-intensity quantitation of the immunoblot, and show the relative amount of Cdc20 normalized to Cdc27. (E) TAME inhibits Cdh1 association with interphase APC. Interphase Xenopus extract was pre-incubated with compounds for 30 min prior to adding recombinant Cdh1 and APC immunoprecipitation. (F) TAME inhibits APC activation by Cdh1. Interphase Xenopus extract was pre-incubated with compound for 30 min prior to adding recombinant Cdh1 and APC immunoprecipitation. The activity of the isolated APC was measured in a reconstituted assay. See also Figure S1.