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Fig. 4. The zinc-finger region of CTCF targets it to the nucleolus. (A) The distribution of the various GFP-CTCF fusion proteins, as described in C, in K562 cells. K562 cells were transfected with the indicated GFP-CTCF truncated variants, cells were fixed 24 hours post-transfection and green fluorescence was analyzed by confocal microscopy. Bar, 5 µm. (B) The distribution of GFP-CTCF fusion proteins in UR61 cells. UR61 cells were transfected with the indicated GFP-CTCF truncated variants (as in C), cells were fixed 24 hours post-transfection and stained with propidium iodide. (a-d) green fluorescence from GFP-CTCF fusion proteins; (e-h) propidium iodide staining labelling nucleoli. Bar, 5 µm. (C) Schematic representation of the full-length and truncated versions of CTCF fused to the C terminus of GFP. Numbers denote amino acid positions. The summary of the nucleolar localization for the different GFP-CTCF fusion proteins in K562, UR61 and HeLa cells is shown. Fusion proteins were present (+) or not (-) in the nucleolus as indicated; (++), strong nucleolar accumulation; (±), weak signal; ND, not determined.
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