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Fig. 5. Nuclear reorganisation within the Hoxd locus. (A)
Four-colour DNA FISH using fosmid probes 3' Hoxd (121N10, yellow),
5' Hoxd (469P2, red) and an MMU2 chromosome paint (green), on DAPI
(blue) counterstained nuclei from undifferentiated (Un) OS25 ES cells and
cells differentiated for 4 and 14 days. Scale bar: 5 µm. (B)
Histograms showing the distribution of 3' Hoxd (121N10, yellow) and
5' Hoxd (469P2, red) hybridisation signals relative to the edge of the
MMU2 CT during differentiation. Arrowheads show the median values for each
probe. (C) Mean position±s.e.m. (µm), relative to the edge of
the MMU2 CT, for all three Hoxd probes during the timecourse of
differentiation. (D) Distribution of interphase separations (d) in
µm, measured between 3' and 5' Hoxd (121N10-469P2) fosmid probe
signals in 2D FISH, during the timecourse of differentiation. Arrowheads show
the median values. In B, C and D, n>100 territories. (E)
Distribution of interphase separations in µm, measured between 121N10 and
860J8 3D FISH signals in undifferentiated ES cells (white bars) and cells
differentiated for 4 days (black bars). Arrowheads show the median values.
(F) Comparison of median interphase distance (d) between signals for
probes 860J8 and 121N10 in undifferentiated cells and after 4 days of
differentiation after 3D (solid line) or 2D (dotted line) FISH.
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