Three-color intranuclear staining for measuring mitosis and apoptosis in cells transfected with a GFP-tagged histone.
Green fluorescent protein (GFP) has become a powerful tool for monitoring the expression of transfected genes by flow cytometry including GFP-tagged histones for tracking chromatin and elucidating histone function. We describe here a method for simultaneous detection of three nucleus-localized signals: a GFP-tagged histone, DNA content and detection of phosphorylated histone H3, which labels mitotic cells. We also demonstrate another application of this method for simultaneous detection of a GFP-tagged histone, DNA content, and cleaved caspase-3.
Rios-Doria
J
J
Risk Assessment, Detection, and Intervention Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
Meléndez
J
J
Monteiro
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AN
D017209:Apoptosis; D053148:Caspase 3 / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D045325:HCT116 Cells; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry*; D006801:Humans; D042581:Intranuclear Space / Q000737:chemistry*; D008938:Mitosis; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D014162:Transfection
D006657:Histones; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D053148:Caspase 3