Figure 6
Cdc7 activity is required for spindle assembly checkpoint. Time-laps images were taken by Olympus LCV110 time-laps microscopy74. M-phase duration was determined for each cell by measuring the time from formation of round cells with condensed chromosomes to separation of daughter cells (cytokinesis) or decondensation of the chromosomes. (a–c) HeLa cells expressing HH2 GFP. Scheme of the experiment along with representative FACS profiles at the end of thymidine block and at 8 hrs after release (a). Movies are shown in Supplementary Movies 5–12. Representative images from the movie are shown in Supplementary Fig. S6a,b. (d,e) AID-mClover-Cdc7 clone #2. The scheme of the experiment along with representative FACS profiles at the end of thymidine block and at 7.5 hrs after release (d). Auxin −, without Auxin; Auxin +, 0.5 µM of Auxin added. Movies are shown in Supplementary Movies 13–18. In (b,c,e), M-phase length for each observed cell is plotted. Bi, 100 nM of AZD1152; Cdc7i, 2 µM of PHA-767491; Cdc7ix, 2 µM of XL413; Noc, 30 ng/ml of Nocodazole; Pacli, 10 nM of paclitaxel. In (b), 10 µM PHA-767491 was also examined.
