Cell viability was determined by trypan blue exclusion assay (Invitrogen) or by flow cytometry of cells stained with the fluorescent DNA intercalator TO-PRO-3 (Invitrogen). The colorimetric 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed as previously described (59 (link)). For cell cycle and death analyses, flow cytometry of cells stained with annexin V and propidium iodide (Invitrogen) was performed using the Guava EasyCyte flowcytometry system (MilliporeSigma), as previously described (66 (link)). Data were analyzed by FCS EXPRESS software (De Novo Software).
Protocol detail
Cell Viability and Cell Death Analyses
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trypan blue exclusion assay
TO-PRO-3
annexin V and propidium iodide
Guava EasyCyte flowcytometry system
FCS EXPRESS software
Annexin v
Assay
Bromide
Cell cycle
Cell viability
Cells
Colorimetric
Dna intercalator
Flowcytometry
Guava
Propidium iodide
Trypan blue
Corresponding Organization :
Other organizations : Medical College of Wisconsin, University of California, San Francisco
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Variable analysis
independent variables
- None explicitly mentioned
- Cell viability
- Cell cycle and death
- None explicitly mentioned
- Positive control: None mentioned
- Negative control: None mentioned
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