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Fig. 2. The HSR is composed of a homogeneous array of transfected plasmid sequences. (A-A'') Metaphase spreads prepared from clone-22 cells were simultaneously hybridized with the DIG-labeled plasmid probe and the biotinylated Alu probe; the hybridized probes were detected with different fluorescent colors. DNA was counterstained with DAPI. (A') The plasmid probe brightly and uniformly stained the HSR (two-headed arrow). (A'') At the HSR, the Alu probe never produced a signal, whereas it brightly labeled highly repetitive Alu sequences on the chromosomes. The signal appeared in bands at the chromosomes, because the Alu sequences were concentrated in the R-band (Matera and Ward, 1992). (B-B'') The metaphase was simultaneously hybridized with the DIG-labeled DM-painting micronuclei probe and the biotin-labeled plasmid probe. (B'') The first probe hybridized with the amplicon already present in the parental COLO 320DM cells, and it painted the DMs in clone-22 cells (compare arrowheads in B and B''), but it never hybridized to the HSR (compare two-headed arrow in B and B''). The DM-derived sequence flanking the HSR is indicated by an arrow (B'').





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