Cell cycle distributions were determined by DNA content assay. HepG2 and Hep3B cells were seeded into six-well plates and treated with the indicated concentrations (0-20 μg/ml) of simvastatin for 48 h. At incubation time, the cells were harvested and fixed in 70% ethanol at 4 °C overnight, and then they were washed with phosphate-buffered saline (PBS) and stained with 20 μg/ml PI at 37 °C for 30 min. The cell populations were analyzed by FAC Sort flow cytometry (BD Biosciences, San Jose, CA, USA).
Fac sort flow cytometry
Manufactured by BD
Sourced in United States
The FAC Sort flow cytometry is a laboratory instrument used for the analysis and sorting of cells or particles in a fluid sample. It utilizes laser-based technology to detect and measure various physical and chemical characteristics of individual cells or particles as they pass through a narrow stream.
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2 protocols using fac sort flow cytometry
1
Cell Cycle Distribution Analysis of HepG2 and Hep3B Cells
2
Mitochondrial Membrane Potential Analysis
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Flow cytometry analysis was done as reported previously15 (link) at Flow Cytometry Core (National Cancer Center). MMP and contents were analyzed by flow cytometry using JC-1, TMRE and MitoTracker as per the manufacturer's instructions. Briefly, cells were dissociated to single cells using trypsin/EDTA and incubated with JC-1 (2 μM ) alone or both TMRE (100 nM ) and 7-AAD (2.5 μg ml−1) or MitoTracker (400 nM ) alone, then analyzed by FACSVerse flow cytometry (BD Biosciences). For cell sorting, dissociated single cells were stained with JC-1 and cells with upper and lower 20% of MMP were sorted using FACSort flow cytometry (BD Biosciences).
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