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Chapters
- 00:05Title
- 01:00Introduction
- 01:34Cell Transfection – Method 1: Nucleofection
- 03:34Cell Transfection – Method 2: Effectene
- 05:43Cell Fusion
- 06:49Fluorescence Microscopy
- 07:44Analysis of Cell-type Specific Protein Localization
- 08:48Conclusion
A flexible and efficient method for the characterization of cell type-specific protein localization and nucleocytoplasmic shuttling is described. This heterokaryon approach uses fluorescently-labeled fusion proteins to image protein localizations after cell fusion. The protocol is amenable to steady-state localizations or more dynamic determinations based on live cell imaging.
Tags
Heterokaryon TechniqueCell Type-specific LocalizationSubcellular TraffickingNucleocytoplasmic ShuttlingProteinsCellular FunctionsNuclear Import/exportTranscriptional RegulationApoptosisCell DivisionDifferentiationTransformationProtein ActivitiesCellular OncogenesisNormal Primary CellsTransformed CellsAltered LocalizationPathwaysDrug TargetingProtein TraffickingShuttling ActivityMammalian CellsHeterokaryon FusionsFluorescence Microscopy
