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Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells

Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells

DOI:
10.3791/62502-v

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09:43 min

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June 13, 2021

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, , ,
1Retrovirus-Host Interactions Laboratory,The Francis Crick Institute, 2Retroviral Replication Laboratory,The Francis Crick Institute, 3Sidney Sussex College, Department of Medicine,University of Cambridge

Chapters

  • 00:05Introduction
  • 00:35Flow Cytometry Analysis
  • 04:18Data Analysis
  • 07:50Results: Analysis of the Restriction Ratio in the Optimal and Suboptimal Data
  • 09:15Conclusion

Summary

Automatic Translation

Automatic Translation

Described here is an established method to determine the extent of HIV-1 restriction by the cellular inhibitory protein SAMHD1. Human myeloid lineage U937 cells are transduced with a SAMHD1 expression vector co-expressing YFP, differentiated and then challenged with HIV-RFP. The level of restriction is determined by flow cytometry analysis.

Tags

Keywords: SAMHD1Flow CytometryU937 CellsHIV InfectionMacrophagesCellular FactorsMulticolor Flow CytometryCytometerFlow RateForward ScatterSide ScatterYFPRFPCompensation Controls

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