Author: Berger, Angela K.; Danthi, Pranav
Title: Reovirus Activates a Caspase-Independent Cell Death Pathway Document date: 2013_5_14
ID: jleccqqx_25
Snippet: Infections and preparation of extracts. ATCC L929 cells were adsorbed with either phosphate-buffered saline (PBS) or reovirus at the indicated MOI at room temperature for 1 h, followed by incubation with medium at 37°C for the indicated time interval. For assessment of NF-B activation, serum-starved cells were infected in the presence of serum-free medium. All inhibitors were added to cells in medium 1 h before virus adsorption and returned with.....
Document: Infections and preparation of extracts. ATCC L929 cells were adsorbed with either phosphate-buffered saline (PBS) or reovirus at the indicated MOI at room temperature for 1 h, followed by incubation with medium at 37°C for the indicated time interval. For assessment of NF-B activation, serum-starved cells were infected in the presence of serum-free medium. All inhibitors were added to cells in medium 1 h before virus adsorption and returned with medium after the 1-h adsorption period. containing a protease inhibitor cocktail (Roche), 500 M dithiothreitol (DTT), and 500 M phenylmethylsulfonyl fluoride (PMSF), followed by centrifugation at 15,000 ϫ g for 10 min to remove debris. Nuclear and cytoplasmic extracts were prepared by hypotonic lysis and high salt extraction, respectively, as previously described (20, 21) .
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