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Supplementary information

From the following article:

The reversibility of mitotic exit in vertebrate cells

Tamara A. Potapova, John R. Daum, Bradley D. Pittman, Joanna R. Hudson, Tara N. Jones, David L. Satinover, P. Todd Stukenberg and Gary J. Gorbsky

Nature 440, 954-958 (13 April 2006)

doi:10.1038/nature04652

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Supplementary Data

This file contains 3 supplementary figures with their legends. The first supplementary figure summarizes the main finding of the paper. The other two contain supplementary data. After the figures is a detailed description of the methods, a short discussion of the effectiveness of Cdk1 inhibitors other than Flavopiridol, and the Supplementary Video Legends.

Supplementary Video 1

This video reveals the effects of the Cdk1 inhibitor Flavopiridol applied to a Xenopus S3 cell in prometaphase, causing premature mitotic exit and cytokinesis. This cell is depicted in Text Fig. 1a.

Supplementary Video 2

This video shows that Flavopiridol induces mitotic exit and cytokinesis in a Xenopus S3 cell arrested at metaphase with the proteasome inhibitor MG132. This cell is depicted in Text Fig. 1b.

Supplementary Video 3

This video shows that Flavopiridol-induced mitotic exit and cytokinesis in a Xenopus S3 cell cultured in the presence of proteasome inhibitor is reversible upon removal of the Flavopiridol. This cell is depicted in Text Fig. 1c. (QuickTime; 3.5MB)

Supplementary Video 4

This video shows a second example of the reversal of mitotic exit and cytokinesis in a Xenopus S3 cell upon removal of Flavopiridol in the presence of proteasome inhibitor.

Supplementary Video 5

This video shows a Xenopus S3 cultured in proteasome inhibitor and induced to go through reversal of mitotic exit by addition then removal of Flavopiridol. The proteasome inhibitor is then washed out and the cell subsequently undergoes normal anaphase and mitotic exit.

Supplementary Video 6

This video shows a Xenopus S3 transiently arrested with the microtubule drug nocodazole then released to proteasome and subsequently induced to undergo mitotic exit and reversal by addition then removal of Flavopiridol. This cell is depicted in Text Fig. 2a.

Supplementary Video 7

This video shows a Hela cell expressing non-degradable cyclin B1. Flavopiridol treatment causes it to exit mitosis and undergo cytokinesis without chromatid separation. Removal of Flavopiridol results in reversal of mitotic exit back to metaphase. This cell is depicted in Text Fig. 3a.

Supplementary Video 8

This video shows a Hela cell expressing non-degradable cyclin B1. Flavopiridol treatment causes it to exit mitosis and undergo cytokinesis after chromatid separation. Removal of Flavopiridol results in reversal of mitotic exit back to an anaphase-like condition. This cell is depicted in Text Fig. 3b.

Supplementary Video 9

This video shows a Hela cell expressing wild type cyclin B1. Flavopiridol treatment causes it to exit mitosis and undergo cytokinesis without chromatid separation. Removal of Flavopiridol does not result in reversal of mitotic exit. This cell is depicted in Text Fig. 3c.

Supplementary Video 10

This video shows that Flavopiridol-induced mitotic exit and cytokinesis in a primary human keratinocyte cell cultured in the presence of proteasome inhibitor is reversible upon removal of the Flavopiridol. This cell is depicted in Text Fig. 4.

Supplementary Video 11

This video shows that R0-31-8220 induces mitotic exit and cytokinesis in a Xenopus S3 cell arrested at metaphase with the proteasome inhibitor MG132. This cell is depicted in Supplementary Fig. 3a.

Supplementary Video 12

This video shows the normal process of mitosis and cytokinesis in a control Xenopus S3 cell stably expressing GFP-alpha-tubulin.

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