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. Author manuscript; available in PMC: 2013 Sep 28.
Published in final edited form as: Mol Cell. 2012 Aug 30;47(6):921–932. doi: 10.1016/j.molcel.2012.07.031

Figure 3. Mad2 and Mad3-Bub3 synergize to inhibit securin ubiquitination and allow autoubiquitination.

Figure 3

(A) Purified unlabeled Cdc20 was pre-incubated with 0.5 μM Mad2 and the indicated Mad3-Bub3 concentration. Reactions were started by the addition of purified 35S-securin, APC/C (5 nM), and E1/E2(Ubc4)/methyl-ubiquitin mix. Similar results were obtained in three independent experiments.

(B) Purified 35S-Cdc20 was pre-incubated with 5 μM Mad2 and the indicated Mad3-Bub3 concentration. Reactions were started by the addition of APC/C (5 nM) and E1/E2(Ubc4)/methyl-ubiquitin mix. Similar results were obtained in two independent experiments.

(C) The data from (A) and (B) were analyzed using Prism, and the zero concentration was plotted on a log scale as 0.01 nM.