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Figure 4


Fig. 4. Representative (A) orangutan (schematic of human chromosome 7, together with the mapping position of large-insert clones, is shown alongside) and (B) Karpas 384 metaphases after FISH with human chromosome-7-specific pools of large-insert clones representing early-replicating (green) and late-replicating (red) loci. (C,D,F,G) 3D maximum intensity projections of nuclei from (C) human fibroblasts, (D) Concolor gibbon lymphoblastoids, (F) orangutan lymphoblastoids and (G) Mel Juso cells hybridized with the early-replicating clone pool labeled green, the late-replicating clone pool in red, chromosome 7 territories blue and DAPI counterstain gray. (E) Human and (H) orangutan z-projected fibroblast nuclei after multicolor 3D-FISH with five BAC clones. The clone color code follows panel K. Scale bar: 5 µm. (I-K,O-Q) Quantitative evaluation of radial probe distributions in the interphase nucleus or the respective chromosome territory, using 3D-RRD or eADS software (n=number of nuclei). (L,M) Mean pair-wise clone distance measurements in human and orangutan fibroblast nuclei from the 3D-FISH experiments illustrated in E,H,K. (N) In both species the mean square interphase distance between two clone loci showed a linear correlation with their genomic distance, indicating a `random walk' 3D conformation of these loci.





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