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Icahn School of Medicine at Mount Sinai, Immunology Institute

3 ARTICLES PUBLISHED IN JoVE

Biology

Eukaryotic Polyribosome Profile Analysis

Anthony M. Esposito¹, Maria Mateyak¹, Dongming He¹, Marcus Lewis¹, Arjun N. Sasikumar¹, Jenna Hutton¹, Paul R. Copeland¹, Terri G. Kinzy¹

¹Department of Molecular Genetics, Microbiology, and Immunology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School

This article describes a protocol for the extraction of translating ribosomes from eukaryotic cells. Once extracted, ribosomes are separated into monosomes and polyribosomes by sucrose gradient fractionation to allow different ribosomal populations to be analyzed. As such, this method is the gold standard for examining the regulation of translation.

Immunology and Infection

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy

Benjamin Dale¹, Gregory P. McNerney², Deanna L. Thompson², Wolfgang Hübner³, Thomas Huser², Benjamin K. Chen¹

¹Division of Infectious Diseases, Department of Medicine, Immunology Institute, Mount Sinai School of Medicine , ²NSF Center for Biophotonics, University of California, Davis, ³Structural and Computational Biology Unit, European Molecular Biology Laboratory

This visualized experiment is a guide for utilizing a fluorescent molecular clone of HIV for live confocal imaging experiments.

Immunology and Infection

A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion

Anthony M. Esposito^1,2, Alexandra Y. Soare¹, Foramben Patel¹, Namita Satija¹, Benjamin K. Chen¹, Talia H. Swartz¹

¹Division of Infectious Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, Immunology Institute, ²Department of Biology, New Jersey City University

We describe a cell-based assay to report on HIV-1 fusion via the expression of green fluorescent protein detectable by flow cytometry or fluorescence microscopy. It can be used to test inhibitors of viral entry (specifically at the fusion step) in cell-free and cell-to-cell infection systems.