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siRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts
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Journal JoVE Génétique
siRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts

siRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts

DOI:
10.3791/53995-v

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09:57 min

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September 20, 2016

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, , ,
1Department of Biological Sciences,University of Quebec in Montreal, 2Department of Pharmacology and Therapeutics,McGill University, 3Department of Clinical Sciences,University of Montreal

Chapitres

  • 00:05Titre
  • 01:00Evaluation of the Purity of Isolated Primary Villous Cytotrophoblasts
  • 03:13siRNA Transfection of Human Primary Villous Cytotrophoblasts
  • 04:55Preparation of Nuclear Extracts from Cultured Cells
  • 05:52Radiolabeling of Single Stranded Oligonucleotide
  • 06:50DNA Binding Reaction
  • 08:07Results: Syncytin-2 Knowckdown and Visualization of Syncytin-2 DNA-protein Complexes
  • 09:14Conclusion

Summary

Traduction automatique

Traduction automatique

This protocol describes a method for efficiently transfecting siRNA in freshly isolated human villous cytotrophoblasts using microporation and identifying DNA-protein complexes in these cells. Transfected cells can be monitored by Western blot and EMSA analyses during the 4-day culture time.

Tags

Keywords: SiRNA TransfectionEMSAHuman Villous CytotrophoblastsPlacenta ResearchGene ExpressionCytotrophoblast DifferentiationRegulatory MechanismsFACS AnalysisCK7 Positive CellsFlow Cytometry

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