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. 2010 Aug 30;107(37):16090–16095. doi: 10.1073/pnas.1005031107

Fig. 1.

Fig. 1.

Effect of the Cdk inhibitor roscovitine on origin firing and replication fork speeds. (A) Labeling protocols for DNA fiber analysis. U2OS cells were pretreated with 25 μM roscovitine (rosc) or an equal volume of DMSO (control) for 1 hour and then pulse labeled with CldU and IdU for 20 minutes each in presence of rosc or DMSO. CldU was detected using a specific primary antibody and a secondary antibody in red. IdU was detected using specific primary antibody and a secondary antibody in green. (B) Representative images of replication tracks from cells treated with DMSO or rosc. (C) Quantification of origin firing in cells treated with DMSO or rosc. First label origins (green-red-green) are shown as percentage of all red (CldU) labeled tracks. (D) Distribution of replication fork speeds in cells treated as in C. (E) Average replication fork speeds in cells treated with DMSO or rosc. Means and standard deviation (S.D.) (bars) of three independent experiments are shown. Values marked with asterisks are significantly different (student’s t-test, ** p < 0.01).