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The intranuclear localization of the Ki-67 reactive antigen was immunocytochemically investigated using flow cytometry. HeLa S3 cells were immunocytochemically stained with the monoclonal antibody, Ki-67, after in situ treatments with various kinds of compounds, namely: HCl; NaCl; RNase; S1 nuclease and DNase I. The only treatment that markedly diminished the immunofluorescence intensity of the cells was exposure to DNase I. Nuclear fluorescence was no longer observed in the cells digested with relatively high concentrations of DNase I. These results suggest that the antigen recognized by Ki-67 is closely associated with DNA, but is not directly associated with either the nuclear matrix or histones.
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