Open AccessCell‐Cycle position and nuclear protein content
Author(s) -
Roti J. L. Roti,
Higashikubo R.,
Blair O. C.,
Uygur N.
Publication year - 1982
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990030205
Subject(s) - cell cycle , nuclear dna , propidium iodide , biology , nuclear protein , microbiology and biotechnology , dna , flow cytometry , staining , pi , fluorescein isothiocyanate , cell , biochemistry , apoptosis , genetics , physics , fluorescence , transcription factor , quantum mechanics , programmed cell death , mitochondrial dna , gene
To determine the change in nuclear protein content as a function of cell cycle position, isolated heLa nuclei were stained for protein with fluorescein isothiocyanate (FITC) and for DNA with propidium iodide (PI) and analyzed by flow cytometry (FCM). The resulting FITC versus PI histogram consisted of four definable regions, a G 1 region characterized by increasing FITC and relatively constant PI (2C DNA content), an S region characterized by increasing PI with relatively constant FITC, a G 2 region characterized by increasing FITC and constant PI (4C DNA content), and a region of G 1 FITC staining with near G 2 PI staining. The relationship between cell cycle position and these regions of the histogram was confirmed by the two following studies: (1) The distribution of labeled nuclei throughout the histogram was observed after [ 14 C]TdR pulse labeling. (2) Exit of cells from G 1 was observed in the histogram after the addition of Colcemid to the HeLa cell cultures. Nuclear protein content did not appear to increase uniformly across the cell cycle (defined by DNA content). Rather, nuclear protein content showed the largest increase during G 1 . Thus, dual parameter FCM analysis based on nuclear DNA and protein content provides a more complete definition of cell cycle position than DNA content alone.