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Fig. 9. Spatial arrangements of GFP-PCNA foci and labelled DNA foci in triple-labelled daughter cells. Equatorial planes of two nuclei labelled and imaged according to the experimental procedure outlined in Fig. 4a are shown. GFP, Cy3 and Cy5 fluorescence and the merged images are shown as indicated (green, GFP; red, Cy3; blue, Cy5; red and green appears yellow; red and blue appears violet; green and blue gives rise to cyan; and a merge of all three colours appears white). (a,b) A daughter nucleus at an earlier stage of S phase during labelling than the mother cell, indicated by relatively numerous GFP-PCNA foci occupying interior positions. The boxed regions are shown enlarged in b. GFP-PCNA foci are adjacent to Cy3-labelled but not to Cy5-labelled DNA foci. (c,d) A nucleus at the transition to a later stage of S-phase than the mother cell during labelling, indicated by the relatively low number of GFP-foci compared to the number of Cy3- or Cy5-labelled foci. The framed regions are shown enlarged in d. (d) GFP-PCNA foci colocalize either with Cy5-labelled foci (upper panels) or sites adjacent to Cy5-labelled foci (lower panels). Bars in a and c, 10 µm; in b, 1 µm; in d, 0.5 µm.
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