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Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms
Journal JoVE
Biologie
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Journal JoVE Biologie
Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms

Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms

DOI:
10.3791/2730-v

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13:40 min

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July 07, 2011

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, , ,
1Medical Biochemistry and Molecular Biology,Saarland University, 2Experimental and Clinical Pharmacology and Toxicology,Saarland University

Chapitres

  • 00:05Titre
  • 02:16Preparation of Stock Solutions
  • 03:03Gene Silencing in HeLa Cells
  • 05:39Complementation of HeLa Cells
  • 07:07Live Cell Calcium Imaging
  • 09:58Representative Calcium Imaging Results
  • 12:58Conclusion

Summary

Traduction automatique

Traduction automatique

The endoplasmic reticulum plays a key role in protein biogenesis and in calcium homeostasis. We have established an experimental system that allows us to address the role of Ca2+ leak channels and to characterize their putative regulatory mechanisms. This system involves siRNA mediated gene silencing and live cell Ca2+ imaging.

Tags

Live Cell Calcium ImagingSiRNA Mediated Gene SilencingCa2+ Leak ChannelsER MembraneRegulatory MechanismsEndoplasmic ReticulumProtein BiogenesisCalcium SignallingHeterotrimeric Sec61 ComplexNascent PolypeptidesPuromycinER Luminal Protein BiPExperimental SystemCoding RegionUntranslated Region (UTR)SEC61A1 GeneComplementation AnalysisIRES-GFP Vector

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