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Usando RNA-interferenza per Indagare sulla risposta immunitaria innata in mouse macrofagi
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JoVE Revista Inmunología y contagio
Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages
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Usando RNA-interferenza per Indagare sulla risposta immunitaria innata in mouse macrofagi

DOI:
10.3791/51306-v

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12:47 min

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November 03, 2014

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, ,
1Integrated Department of Immunology and Integrated Center for Genes, Environment, and Health,National Jewish Health and University of Colorado School of Medicine

Capítulos

  • 00:05Título
  • 01:26Maintenance of Microphage Cell Lines
  • 02:04Programming the 96-well Shuttle System for Transfection
  • 03:14Preparing the Reagents and Macrophages for Transfection
  • 05:27Nucleofection of Macrophages with siRNA
  • 07:45Recovery and Plating of Macrophages
  • 09:35Stimulation of Macrophages with LPS and Monitoring the Induced Innate Immune Response, Gene Knockdown and Viability in Macrophages
  • 10:58Results: Monitoring the Innate Immune Response
  • 12:11Conclusion

Summary

Traducción Automática

Traducción Automática

In this protocol, we describe methods to efficiently transfect murine macrophage cell lines with siRNAs using the Amaxa Nucleofector 96-well Shuttle System, stimulate the macrophages with lipopolysaccharide, and monitor the effects on inflammatory cytokine production.

Tags

RNA InterferenceMacrophageInnate Immune ResponseGene KnockdownSiRNA TransfectionAmaxa Nucleofector96-well FormatLipopolysaccharideCytokine Production

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