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

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

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

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).

ISSN: 1743-422X