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The ability to transiently express a gene of interest while simultaneously monitoring cell cycle progression in a controlled environment will facilitate and better quantify many experiments in yeast cell biology.
The effects of such treatments on the induction of cellular apoptosis have been evaluated by monitoring cell cycle alterations and caspase involvement.
To address that question in a more systematic and quantitative manner than immunostaining detection, we developed spheroid models expressing Fucci (Fluorescence Ubiquitination Cell Cycle Indicators) tools that allow monitoring cell cycle position of a cell [ 19].
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Our approach was to express the phosphorylation site mutant CDK1AF, which short-circuits the feedback loops, in HeLa cells, and to monitor cell cycle progression by live cell fluorescence microscopy.
We then monitored cell cycle progression by flow cytometry.
Then we monitored cell cycle recovery in response to MMC in both U2OS and HeLa cells.
One way to monitor cell cycle progression is with populations of dividing cells.
To investigate the effect of these knockdowns on cell cycle progression, we monitored cell cycle distribution by flow cytometry.
FACS analysis was used as an additional means to quantitate apoptotic, dead and live cell fractions, as well as to monitor cell cycle.
To investigate if the different effect of BBR derivatives on the two cell lines could be ascribed to a diverse impact on cell cycle, we monitored cell cycle distribution by flow cytometry.
Live cell imaging of fluorophore-tagged cell cycle markers has been used to monitor cell cycle progression in unsynchronized cell populations, but it requires stable protein overexpression and is limited in the use of additional markers.
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