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Table 1 Quantitative assessment of the positions of integrated operator sequences and control genomic sites relative to PML or Sp100

From: Nuclear domain 10-associated proteins recognize and segregate intranuclear DNA/protein complexes to negate gene expression

DNA locus Cell line Colocali-zation Side by side orpartial overlap No association Number of nuclei evaluated
large operator insert w/o repressor QT10 HEp-2-Op256 0% 26% 74% 1000
large operator insert + GFP-repressor QT10 HEp-2-Op256 100% 0% 0% 1000
small operator insert w/o repressor QT5 HEp-2-Op128 0% 12% 87% 1000
small operator insert + GFP-repressor QT5 HEp-2-Op128 72% 0% 28% 1000
U2 human fibroblasts 0% 17% 83% 960
D1Z2 human fibroblasts 0% 11% 89% 1280
β-actin human fibroblasts 0% 22% 78% 940
collagen Iα1 human fibroblasts 0% 24% 76% 1070
  1. Association of ND10 with integrated repetitive sequences (lac operator, with or without binding GFP-repressor), endogenous repetitive sequences (U2, D1Z2), and endogenous non-repetitive genes (β-actin, collagen Iα1). DNA loci were visualized either by FISH or via the binding GFP-repressor, and cells were simultaneously stained for PML or Sp100 to assess the percentage of loci that associate with these proteins (side-by-side and/or overlapping). Loci were counted as colocalizing if the site was totally covered by PML or Sp100, as side-by-side if touching or partially overlapping, and as not associated if a clear space could be discerned between the two structures. Genomic sites consisting of repetitive DNA (U2, D1Z2) do not show a higher association rate with ND10 compared to that seen with single copy genes (β-actin, collagen Iα1), and neither one colocalizes with PML or Sp100 (QT10: HEp-2-Op256; QT5: HEp-2-Op128).