Selected article for: "cell seeding and protein expression"
Author: Griffiths, Samantha J.; Koegl, Manfred; Boutell, Chris; Zenner, Helen L.; Crump, Colin M.; Pica, Francesca; Gonzalez, Orland; Friedel, Caroline C.; Barry, Gerald; Martin, Kim; Craigon, Marie H.; Chen, Rui; Kaza, Lakshmi N.; Fossum, Even; Fazakerley, John K.; Efstathiou, Stacey; Volpi, Antonio; Zimmer, Ralf; Ghazal, Peter; Haas, Jürgen
Title: A Systematic Analysis of Host Factors Reveals a Med23-Interferon-? Regulatory Axis against Herpes Simplex Virus Type 1 Replication
  • Document date: 2013_8_8
    • ID: 0lyt8gfq_62
    Snippet: Induction of IFN-l by Med23 was determined in a range of cell types by seeding cells in 96-well plates to be ,80% confluent the next day. Cells were transfected in duplicates with 100 ng pCR3 or pCR3-Med23 using Lipofectamine LTX with Plus reagent (Invitrogen), in antibiotic-free medium. IFN-l levels quantified 96-120 h post-transfection. The synergistic effect of Med23 and IRFs on IFN-l induction was determined by co-transfection of pCR3 or pCR3.....
    Document: Induction of IFN-l by Med23 was determined in a range of cell types by seeding cells in 96-well plates to be ,80% confluent the next day. Cells were transfected in duplicates with 100 ng pCR3 or pCR3-Med23 using Lipofectamine LTX with Plus reagent (Invitrogen), in antibiotic-free medium. IFN-l levels quantified 96-120 h post-transfection. The synergistic effect of Med23 and IRFs on IFN-l induction was determined by co-transfection of pCR3 or pCR3-Med23 (50 ng) with pCR3-IRF7 (50 ng) in A549 cells, with Lipofectamine LTX with Plus reagent. The effect of Med23 depletion on IFN-l induction was determined by transfection of A549 cells in 96-well plates with 50 nM RSCF or Med23 siRNA. IFN-l was quantified 120 h post-transfection. IFN-l protein expression was quantified in supernatants using a commercial IFN-l DuoSet ELISA kit (R and D Systems).

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