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Fig. 6. Analysis of Cy3 and GFP-PCNA colocalization patterns. (a) A scheme summarizing the spatiotemporal dynamics observed. During the temporal progression of S phase (indicated by black arrows) DNA foci (red) stably occupy their nuclear positions, whereas GFP-PCNA foci appear at adjacent sites. A transition stage is observed during which GFP foci at the previously active sites cease, whereas GFP foci at adjacent new sites increase (compare with Fig. 3a and b, 58 minutes and 82 minutes). Colocalization with labelled DNA (middle) is only observed during a restricted period (compare Figs 2 and 5). This scheme illustrates the behaviour of the majority of replication sites and does not take into account intranuclear heterogeneity, which might occur when the replication programme is not exactly reproduced at all sites. (b-d) Quantitative analysis of colocalization between Cy3 and GFP-PCNA foci in daughter cells. 21872 Cy3-labelled and 17401 GFP-labelled foci were evaluated. Red and yellow bars represent the percentage of Cy3-labelled foci colocalizing (yellow) or not colocalizing (red) with GFP-PCNA labelled foci. Green bars represent the percentage of GFP-labelled foci not colocalizing with Cy3-labelled foci. Individual nuclei display different degrees of colocalization. (b) In class I nuclei on average only about 15% of Cy3-labelled foci colocalize with GFP-labelled foci. (c) In class II nuclei about half of the Cy3-labelled foci colocalize with GFP-PCNA foci. (d) In class III nuclei on average about 85% of Cy3-labelled foci colocalize with GFP-labelled foci.
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