Figure 2. Dormant origins activated within existing replication factories in response to replicative stress. U2OS cells were synchronized in early S phase and treated with HU for 2 h as in Fig. 1. Cells were pulsed with Cy3-dUTP for 30 min before fixing. (a and b) Representative images of control (a) and 200 µM HU-treated cells (b). (c–f) The intensity of individual Cy3-dUTP foci was measured and averaged in each cell. (c and e, closed squares) Analysis of 50 cells under each condition gave rise to the overall mean foci intensity with SEM (error bars). (c and e, open squares) The replication fork speed of the samples was measured by DNA fiber analysis. The ratio of mean Cy3-dUTP focus intensity to fork speed (d and f) indicates the relative number of forks within each replication focus.
Figure 2.

Dormant origins activated within existing replication factories in response to replicative stress. U2OS cells were synchronized in early S phase and treated with HU for 2 h as in Fig. 1. Cells were pulsed with Cy3-dUTP for 30 min before fixing. (a and b) Representative images of control (a) and 200 µM HU-treated cells (b). (c–f) The intensity of individual Cy3-dUTP foci was measured and averaged in each cell. (c and e, closed squares) Analysis of 50 cells under each condition gave rise to the overall mean foci intensity with SEM (error bars). (c and e, open squares) The replication fork speed of the samples was measured by DNA fiber analysis. The ratio of mean Cy3-dUTP focus intensity to fork speed (d and f) indicates the relative number of forks within each replication focus.

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