Author: Bederka, Lydia H.; Bonhomme, Cyrille J.; Ling, Emily L.; Buchmeier, Michael J.
Title: Arenavirus Stable Signal Peptide Is the Keystone Subunit for Glycoprotein Complex Organization Document date: 2014_10_28
ID: wbh06gzb_34
Snippet: Fusion assay. DBT cells were plated onto flame-sterilized glass coverslips and transfected using Lipofectamine 2000 (Life Technologies). After 48 h of incubation, cells were washed with 1Ï« PBS and incubated with pH 5-adjusted DMEM growth medium for 1 h before returning to pH 7.4 DMEM maintenance medium for an additional 1 h. Cells were fixed with 3.7% paraformaldehyde and extensively washed with 1Ï« PBS, and coverslips were mounted with DAPI-con.....
Document: Fusion assay. DBT cells were plated onto flame-sterilized glass coverslips and transfected using Lipofectamine 2000 (Life Technologies). After 48 h of incubation, cells were washed with 1Ï« PBS and incubated with pH 5-adjusted DMEM growth medium for 1 h before returning to pH 7.4 DMEM maintenance medium for an additional 1 h. Cells were fixed with 3.7% paraformaldehyde and extensively washed with 1Ï« PBS, and coverslips were mounted with DAPI-containing mounting medium. Images were acquired using the Nikon Eclipse Ti microscope. The fusion index was determined by counting the number of cells versus the total nuclei, as previously described (29) .
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