Technical Reference #495

495.

Visualization Of Retroviral Replication In Living Cells Reveals Budding Into Multivesicular Bodies Nathan M. Sherer; Maik J. Lehmann; Luisa F. Jimenez-Soto; Alyssa Ingmundson; Stacy M. Horner; Gregor Cicchetti; Philip G. Allen; Marc Pypaert; James M. Cunningham and Walther Mothes, Yale University, Traffic, 4(495), (2003)
Link To Paper

Abstract:
Retroviral assembly and budding is driven by the Gagpolyprotein and requires the host-derived vacuolar proteinsorting (vps) machinery. With the exception ofhuman immunodeficiency virus (HIV)-infected macrophagescurrent models predict that the vps

Keywords:
human immunodeficiency virus (HIV); imaging of viral replication; multivesicular body (MVB); murine leukemia virus (MLV); retroviral budding and assembly

Materials & Methods:
For live confocal imaging cells were transferred 10–16 h post transfection to poly L-lysine coated 35-mm glass-bottom plates (MatTek Ashland MA USA). At 24–48 h post transfection medium was replaced with DMEM (10% FBS) supplemented with 10mM HEPE

Microscopic Technique
Confocal Microscopy

Cell Type(s)
HEK-293