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. Author manuscript; available in PMC: 2012 Apr 13.
Published in final edited form as: Dev Cell. 2010 Apr 20;18(4):533–543. doi: 10.1016/j.devcel.2010.02.013

Figure 5. Centrosome separation is not a consistent marker for CyclinB1 activation.

Figure 5

A&C. Asynchronous HeLa cells co-expressing the CyclinB1-Cdk1 biosensor and mCherry-α–tubulin were followed by time-lapse fluorescence microscopy at 1 image/2 min. The FRET quantification curves of 3 different cells are displayed and the beginning of centrosome migration for each cell is indicated by a vertical dashed line (C). B&D. Synchronized RPE cells co-expressing the CyclinB1-Cdk1 sensor and mCherry-α-tubulin were followed by time-lapse fluorescence microscopy at 1 image/2 min. The FRET quantification curves of 3 different cells are displayed. The beginning of centrosome migration for each cell is indicated by a vertical dashed line (D). E. HeLa cells co-expressing the CyclinB1-Cdk1 biosensor and mCherry- α-tubulin were synchronized in S phase by a thymidine block and release regime and assayed by time-lapse fluorescence and DIC microscopy. Upper panel: In the cell displayed the centrosomes separated >50 min before NEBD. Bottom panel: time between beginning of FRET increase and centrosome separation in typical experiments using asynchronous (n=14) or synchronized (n=12) HeLa cells. Note that centrosome separation begins at random during G2 phase in synchronised HeLa cells.