Cell-line generation and maintenance

The A549 cell-line (ECACC) and derivatives (Appendix Table A.1) were maintained in DMEM with 10% fetal bovine serum (FBS) (DMEM-10). A549 reporter cell-line derivatives expressing HCV NS3-4A, RSV NS1/NS2 or PIV5 V were generated using a self-inactivating lentiviral constitutive expression system (Demaison et al., 2002 (link)). A549 reporter cell-line derivatives expressing CMV IE1 and corresponding negative controls (luciferase (Luc) and IE1dl410-420 (Harwardt et al., 2016 (link))) were generated using the Lenti-X Tet-One inducible expression system (Clontech). Lentiviruses were generated in 293T cells (ECACC), used to transduce the appropriate reporter cell-line and maintained under antibiotic selection. A Hep2 cell-line derivative constitutively expressing PIV2 V (Precious et al., 2005 (link)) or BVDV Npro (Hilton et al., 2006 (link)), MRC-5 cells (ATCC), Vero cells (ECACC) and STAT2-deficient primary skin fibroblasts (Hambleton et al., 2013 (link)) were maintained in DMEM-10.

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Vasou A., Paulus C., Narloch J., Gage Z.O., Rameix-Welti M.A., Eléouët J.F., Nevels M., Randall R.E, & Adamson C.S. (2018). Modular cell-based platform for high throughput identification of compounds that inhibit a viral interferon antagonist of choice. Antiviral Research, 150, 79-92.

Publication 2018

Lenti-X Tet-One inducible expression system MRC-5 cells

293t cells A549 cell line Antibiotic Cell line Cells Cmv ie1 Derivatives Fetal bovine serum Fibroblasts Hcv ns3 4a Lentiviruses Luciferase Skin Stat2 Vero cells

Corresponding Organization :

Other organizations : University of St Andrews, Assistance Publique – Hôpitaux de Paris, Virologie et Immunologie Moléculaires

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Variable analysis

independent variables

Expressing HCV NS3-4A, RSV NS1/NS2 or PIV5 V in A549 reporter cell-line derivatives
Expressing CMV IE1 and corresponding negative controls (luciferase (Luc) and IE1dl410-420) in A549 reporter cell-line derivatives

dependent variables

Not explicitly mentioned

control variables

DMEM with 10% fetal bovine serum (FBS) (DMEM-10) used to maintain cell lines
Antibiotic selection used to maintain transduced cell lines
Hep2 cell-line derivative constitutively expressing PIV2 V or BVDV Npro
MRC-5 cells, Vero cells, and STAT2-deficient primary skin fibroblasts maintained in DMEM-10

positive controls

Hep2 cell-line derivative constitutively expressing PIV2 V
Hep2 cell-line derivative constitutively expressing BVDV Npro

negative controls

A549 reporter cell-line derivatives expressing luciferase (Luc)
A549 reporter cell-line derivatives expressing IE1dl410-420

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