Laser Flow Cytofluorometric Analysis of Htc Cells Synchronized With Hydroxyurea, Nocodazole and Aphidicolin

Miller-Faurès, A.; Michel, Nico; Aguilera, Andrés; Blave, A.; Miller, A. O. A.

doi:10.1111/j.1365-2184.1981.tb00556.x

Cited by 4 publications

(2 citation statements)

References 16 publications

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“…The percentage of cells in G0/G1, S and G2+M was determined by DNA staining with propidium iodide (15). Cells were fixed in 70% ethanol and incubated overnight at 4 ° C. Cells were resuspended in MEM containing 2% newborn calf serum, 500 U/ml RNase A and 0.1% sodium azide and incubated at 37 ° C for 1 h. An equal volume of 50 ~tg/ml propidium iodide in 1.12% sodium citrate, pH 8.4 was added and cells were incubated at room temperature for 30 rain.…”

Section: Cell Cycle Analysismentioning

confidence: 99%

Cell cycle progression is associated with distinct patterns of phosphorylation of Op18

Strahler

Lamb

Ungar

et al. 1992

Biochemical and Biophysical Research Communications

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Op18 is a highly conserved major cytosolic phosphoprotein which has been implicated in signal transduction in a wide variety of cell types. Freshly isolated peripheral blood lymphocytes (PBL) constitutively express low levels of mostly unphosphorylated Op18. Following mitogenic stimulation of PBL, Op18 synthesis is induced at a time when cells are entering S-phase. In this study we have characterized Op18 phosphorylation during progression of freshly isolated PBL through the cell cycle. Transition from G0 to G1 following activation with OKT3 was associated with an increase in a phosphorylated form designated Op18c. Progression of cells through G1 into S resulted in an increase in phosphorylated Op18 forms, designated Op18a and Op18b, which paralleled new Op18 synthesis. Transition of cells into G2 + M resulted in the appearance of the more acidic phosphorylated forms Op18d and Op18e. Calphostin C, a specific inhibitor of protein kinase C, dramatically decreased all forms of phosphorylated Op18 in OKT3 treated Jurkat cells. Our results suggest that Op18 phosphorylation is mediated in part by PKC activation as well as by other kinases yielding different phosphorylated forms at specific stages of the cell cycle.

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Section: Cell Cycle Analysismentioning

confidence: 99%

Cell cycle progression is associated with distinct patterns of phosphorylation of Op18

Strahler

Lamb

Ungar

et al. 1992

Biochemical and Biophysical Research Communications

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“…If cells are arrested for extended periods in mitosis or with higher concentrations of drug, mitotic spindles reform less efficiently with a large percentage of cells incapable of returning to interphase (Zieve et al 1980). Prolonged exposures to nocodazole also cause irreversible polyploidization and the formation of micronuclei when some cells exit mitosis without dividing (Miller-Faurè s et al 1981;Nü sse and Egner 1984). Hence, short exposures to small concentrations of nocodazole would seem in theory to be the optimal way, but it can provide only small numbers of mitotic cells.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Mammalian Cell-free Systems of Nuclear Disassembly and Assembly

Vaillant

Paulin−Levasseur

2007

J Histochem Cytochem.

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Mammalian cell-free systems are very useful for the biochemical and structural study of nuclear disassembly and assembly. Through experimental manipulations, the role of specific proteins in these processes can be studied. Recently, we intended to examine the involvement of integral and peripheral inner nuclear membrane proteins in nuclear disassembly and assembly. However, we could not achieve proper disassembly when isolated interphase HeLa nuclei were exposed to mitotic soluble extracts obtained from the same cell line and containing cyclin B1. Homogenates of synchronized mitotic HeLa cells left to reassemble their nuclei generated incomplete nuclear envelopes on chromatin masses. Digitonin-permeabilized mitotic cells also assembled incomplete nuclei, generating a lot of cytoplasmic inclusions of inner nuclear membrane proteins as an intermediate. These results were therefore used as a basis for a critical evaluation of mammalian cell-free systems. We present here evidence that cell synchronization itself can interfere with the progress of nuclear assembly, possibly by causing aberrant nuclear disassembly and/or by inducing the formation of an abnormal number of mitotic spindles.

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Dielectric spectroscopy of mammalian cells

Degouys

Cerckel²,

Garcia³

et al. 1993

Cytotechnology

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Low-frequency dielectric spectroscopy has been used in situ, i.e. while the cells are still attached to their microsupport, to monitor the changes of biomass accompanying the growth of anchorage-dependent cells. This method, when compared to Aperture Impedance Pulse Spectroscopy (also called electronic sizing), is characterized by a somewhat lower degree of resolution. Suggestions are made on how to determine the capacitance of the spent growth medium alone, still keeping the probe inserted in the bioreactor. This will make dielectric spectroscopy the first truly in situ, on-line, in real time, non-invasive measure of the biomass.

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Laser Flow Cytofluorometric Analysis of Htc Cells Synchronized With Hydroxyurea, Nocodazole and Aphidicolin

Cited by 4 publications

References 16 publications

Cell cycle progression is associated with distinct patterns of phosphorylation of Op18

Cell cycle progression is associated with distinct patterns of phosphorylation of Op18

Evaluation of Mammalian Cell-free Systems of Nuclear Disassembly and Assembly

Dielectric spectroscopy of mammalian cells

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