Article Title: Identification of an endogenous retroviral envelope gene with fusogenic activity and placenta-specific expression in the rabbit: a new "syncytin" in a third order of mammals
Figure Lengend Snippet: Fusion assay between ASCT2 -transduced and syncytin-Ory1 -transduced co-cultured cells demonstrates that ASCT2 is the syncytin-Ory1 receptor . Left panel: Cell-cell fusion was assayed upon independent transfections of a set of A23 cells with an empty vector (none) or an expression vector for either the syncytin-Ory1, syncytin-1 or syncytin-2 protein together with an nls- LacZ gene-expression vector, and another set of A23 cells with an expression vector for the syncytin-1 receptor ASCT2, the syncytin-2 receptor MFSD2 [ 13 ] or an empty vector (none). One day after transfection, cells were resuspended and pairs of transfected cells from each set were cocultured for 1-2 days, fixed and X-Gal stained. Right panel: Syncytia can be easily detected (arrows) for the syncytin-Ory1/ASCT2, syncytin-1/ASCT2 and syncytin-2/MFSD2 pairs, with only mononucleated cells visible in the other cases. Abbreviations: syn-Ory1, syncytin-Ory1; syn1, syncytin-1; syn2, syncytin-2.
Article Snippet: For the self-fusion assay, cells seeded at 104 - 5 × 104 cells per well in 24-well plates were transfected by using the Lipofectamine kit (Invitrogen) with 0.2 μg of either the syncytin-Ory1 expressing or an empty vector, supplemented with 0.2 μg of a LacZ-expression vector (pCMV-β, Clontech).
Techniques: Single Vesicle Fusion Assay, Cell Culture, Transfection, Plasmid Preparation, Expressing, Staining