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Immunology and Infection

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JoVE Journal Immunology and Infection
Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells
 

Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells

Article DOI: 10.3791/59697-v • 06:21 min • August 27th, 2019
August 27th, 2019 •
1, 2
1Norfolk Academy, 2Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale School of Medicine

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We describe a protocol utilizing fluorescence in situ hybridization (FISH) to visualize multiple herpesviral RNAs within lytically infected human cells, either in suspension or adherent. This protocol includes quantification of fluorescence producing a nucleocytoplasmic ratio and can be extended for simultaneous visualization of host and viral proteins with immunofluorescence (IF).

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Keywords: Quantitative FISH Immunofluorescence KSHV Host-virus Interactions Viral Gene Products RNA Protein Biochemical Assays Cell Culture Fixation Permeabilization Blocking Primary Antibody Secondary Antibody Fluorescent Labeling Hybridization Oligonucleotides Denaturation
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