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. Author manuscript; available in PMC: 2012 Oct 1.
Published in final edited form as: Nat Struct Mol Biol. 2012 Mar 11;19(4):387–394. doi: 10.1038/nsmb.2245

Figure 6.

Figure 6

Checkpoint activation by mitotic telomere deprotection. (a) Immunofluorescence of synchronized IMR90 cells treated with 100 ng ml-1 colcemid 6 h post release, cultured for 18 h before collection by shake-off. The cells were replated and fixated at the indicated times. A mitotic cell 10 h post-release without colcemid serves as the negative control. (b) Schematic of the timing of the experiment in panel c. (c) Western analysis of asynchronous IMR90 cells or IMR90 cells synchronized and collected as in panel b. Antibodies used are indicated on the left. (d) Schematic of the timing of the experiment in panel e. (e) Quantification of BrdU-positive cells in asynchronous IMR90 and synchronized IMR90 and IMR90 E6E7 populations treated as in panel d. BrdU incorporation and DNA content was determined by FACS analysis. The mean and standard deviation of three independent experiments (20,000 cells analyzed per time point) is shown. Scale bar, 10 µm.