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Metoder til klassificering af cytoplasmatiske foci som spædbørn
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JoVE Journal Biology
Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules

Metoder til klassificering af cytoplasmatiske foci som spædbørn

DOI:
10.3791/55656-v

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09:33 min

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May 12, 2017

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, , , , ,
1Division of Rheumatology, Immunology, and Allergy,Brigham and Women’s Hospital, 2Department of Medicine,Harvard Medical School, 3Department of Histology and Embryology,Poznan University of Medical Sciences, 4The Broad Institute of Harvard and M.I.T.

Chapters

  • 00:05Title
  • 00:52Stressing Cells
  • 02:19Cell Fixation and Staining
  • 05:23Fluorescence In Situ Hybridization
  • 07:43Results: Stress Granules Formation in Sodium Arsenite- and Vinorelbine-treated Cells
  • 08:46Conclusion

Summary

Automatic Translation

Automatic Translation

Stress Granuler (SG'er) er ikke-membraniske cytoplasmatiske strukturer, der dannes i celler udsat for en række stress. SG'er indeholder mRNA'er, RNA-bindende proteiner, små ribosomale underenheder, translationsrelaterede faktorer og forskellige cellesignalproteiner. Denne protokol beskriver en workflow, der bruger flere eksperimentelle metoder til at detektere, karakterisere og kvantificere bona fide SG'er.

Tags

Cytoplasmic FociStress GranulesMammalian CellsU-2 OS CellsSodium ArseniteVinorelbineCycloheximidePuromycinImmunofluorescenceCell FixationCell Permeabilization

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