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. Author manuscript; available in PMC: 2016 Mar 16.
Published in final edited form as: Curr Biol. 2015 Mar 5;25(6):811–816. doi: 10.1016/j.cub.2015.01.053

Figure 2. Full Stabilization of Cyclin B3 Requires Both the SAC and Early Cyclins.

Figure 2

(A) Schematic of the experiments showing the approximate stage of each injection.

(B and C) Colchicine injection induced mitotic arrest in control, but not in Cyclin A+B knockdown, embryos. Mitotic chromosomes were visualized by H2AvD-RFP (red), and nuclear envelope was visualized by injection of fluorescently labeled wheat germ agglutinin (WGA) (green). Compare based on indicated timing (min:s) not alignment.

(D–F) Characterization of Cyclin B3 degradation under different conditions. Cyclin B3-GFP (green) was stabilized by colchicine in wild-type, but not in Cyclin A+B-depleted, embryos. APC/C inhibitor UbcH10C114S blocked this destruction. Mitotic chromosomes were visualized by H2AvD-RFP (red). Note that in (E) chromosomes exited from mitosis after Cyclin B3-GFP degradation. Scale bar, 5 mμ.

(G) Quantitation of Cyclin B3-GFP fluorescent intensity after colchicine injection. Total fluorescence of each frame was measured, normalized, and plotted against time (minutes into mitosis). Cyclin B3-GFP was similarly stabilized by injection of recombinant Cyclin B proteins and a known inhibitor of the APC/C, UbcH10C114S.

See also Figures S1 and S2 and Movie S2.