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In Vitro Polymerization of F-actin on Early Endosomes
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Biochemistry
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JoVE Journal Biochemistry
In Vitro Polymerization of F-actin on Early Endosomes

In Vitro Polymerization of F-actin on Early Endosomes

DOI:
10.3791/55829-v

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12:15 min

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August 28, 2017

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, , , ,
1Department of Biochemistry,University of Geneva, 2Department of Fundamental Microbiology,University of Lausanne

Chapters

  • 00:05Title
  • 00:53Endosomal Fraction Preparation
  • 06:10In Tube Assay for Endosome-dependent Actin Polymerization
  • 08:11In Chamber Assay for Endosome-dependent Actin Polymerization
  • 09:29Results: F-actin Polymerization on Early Endosomes
  • 11:09Conclusion

Summary

Automatic Translation

Automatic Translation

Early endosome functions depend on F-actin polymerization. Here, we describe a microscopy-based in vitro assay that reconstitutes the nucleation and polymerization of F-actin on early endosomal membranes in test tubes, thus rendering this complex series of reactions amenable to biochemical and genetic manipulations.

Tags

Keywords: In VitroF-actinPolymerizationEarly EndosomesMembrane TraffickingActin FieldsAcidic PathwayNucleationReconstitutionHeLa CellsHomogenization BufferCell FractionationPhase Contrast Microscopy

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