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Fig. 2. Spatial relationship of the Mash1 locus to centromeric heterochromatin and the nuclear periphery. (A) By 3D FISH we observed that the Mash1 locus (green) is rarely associated with centromeric heterochromatin ({gamma}-satellite probe is red) in either ES or neural cells. Numbers in brackets are FISH signal scores. (B) Frequency (mean ± s.d.) of FISH signals scored as peripheral (≥80% of nuclear radius) in undifferentiated OS25 ES cells and differentiated OS25 neural cells. Numbers in bars are total fish signals scored. (C) Representative 2D FISH result showing Mash1 locus (green) in OS25 ES and neural cells. Nuclei were counterstained with DAPI (blue). (D) Representative 3D FISH result showing Mash1 locus (green) in individual z slices of DAPI-stained ES and neural nuclei. Frequency of perinuclear Mash1 signals scored in 3D analysis is indicated. Numbers in brackets represent the total FISH signals scored. Bar, 5 µm.