Author: Dejnirattisai, Wanwisa; Webb, Andrew I.; Chan, Vera; Jumnainsong, Amonrat; Davidson, Andrew; Mongkolsapaya, Juthathip; Screaton, Gavin
Title: Lectin Switching During Dengue Virus Infection Document date: 2011_6_15
ID: qos9vu3r_11
Snippet: Cells were infected with DENV at a multiplicity of infection (MOI) of 1 for 24 h. Cells were washed twice in FACS wash (FW; Phosphate-Buffered Saline [PBS] containing .5% Bovine Serum Albumin, 2% FBS, 1% Human Serum, .01% NaN 3 ). Cells were fixed with 4% paraformaldehyde/PBS for 10 minutes and permeabilized with .5% saponin in FW for 10 minutes. Cells were then incubated with anti-NS1 mAb or with anti-E mAb followed by anti-mouse IgG/PE (DakoCyt.....
Document: Cells were infected with DENV at a multiplicity of infection (MOI) of 1 for 24 h. Cells were washed twice in FACS wash (FW; Phosphate-Buffered Saline [PBS] containing .5% Bovine Serum Albumin, 2% FBS, 1% Human Serum, .01% NaN 3 ). Cells were fixed with 4% paraformaldehyde/PBS for 10 minutes and permeabilized with .5% saponin in FW for 10 minutes. Cells were then incubated with anti-NS1 mAb or with anti-E mAb followed by anti-mouse IgG/PE (DakoCytomation) in .5% saponin/FW. Cells were resuspended in FW and analyzed by FACScan (Becton Dickinson). Data were analyzed by using Flowjo software (Tree Star). 3T3 and 293T were incubated with 20 U/mL heparin for 20 minutes at room temperature before being infected with DENV.
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