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. 2011 Aug 29;31(15):1975–1987. doi: 10.1038/onc.2011.372

Figure 2.

Figure 2

Cdc20 induces RASSF1A degradation. The protein level of RASSF1A fluctuates across the cell cycle progression. HeLa cells were synchronized by double thymidine block followed by releasing into medium without thymidine (a), double thymidine block followed by releasing into nocodazole (b, left panel), or nocodazole block and release (b, right panel). Cells collected at indicated time points were analyzed by immunoblotting. Similar to Cyclin A, RASSF1A was degraded at 8 h after releasing from thymidine block in the presence or absence of nocodazole. (c) Ectopic expression of Cdc20 downregulates RASSF1A. HeLa cells were transfected with vectors encoding FLAG-Cdc20(7A) or FLAG-Cdh1 and synchronized by double thymidine block, followed by releasing into nocodazole. The level of RASSF1A at indicated time points were analyzed by immunoblotting. (d) Knocking down of Cdc20 by siRNA stabilizes RASSF1A during early mitosis. HeLa cells transfected with indicated siRNA were synchronized and analyzed as in (c).