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Methoden om Cytoplasmatische Foci te classificeren als Mammalian Stress Granules
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Biologia
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JoVE Journal Biologia
Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules
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Methoden om Cytoplasmatische Foci te classificeren als Mammalian Stress Granules

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.

Capitoli

  • 00:05Titolo
  • 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

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Stress Granules (SG's) zijn niet-membraan cytoplasmatische structuren die in cellen vormen die blootstaan ​​aan een verscheidenheid aan stressen. SG's bevatten mRNA's, RNA-bindende eiwitten, kleine ribosomale subeenheden, vertaalgerelateerde factoren en diverse cel signalerende eiwitten. Dit protocol beschrijft een workflow die gebruik maakt van verschillende experimentele benaderingen om bona fide SG's te detecteren, karakteriseren en kwantificeren.

Tags

Cytoplasmic FociStress GranulesMammalian CellsU-2 OS CellsSodium ArseniteVinorelbineCycloheximidePuromycinImmunofluorescenceCell FixationCell Permeabilization

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