Human epithelial cell HEK-293T (ATCC® CRL-3216™), simian epithelial cell Vero E6 (ATCC® CRL-1586™), human astrocytoma cell U-87MG (ATCC® HTB-14™), and human neuroblastoma cell SH-SY5Y (ATCC® CRL-2266™) were provided by CelluloNet (SFR-Biosciences, Lyon, France). HEK-293T, Vero E6, and U-87MG cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; GlutaMAX, high glucose, Gibco™, Thermo Fisher Scientific, Carlsbad, CA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco™). SH-SY5Y cells were grown in DMEM/nutrient mixture F-12 (DMEM/F-12, Ham 1:1, GlutaMAX, Gibco™) supplemented with 2% heat-inactivated FBS. Cell cultures were maintained at 37 °C and 5% CO2. For the production of recombinant sNS1 protein, geneticin™-G418 (500 μg/mL, Gibco™) was added to the culture medium of HEK-293T cells 3 days before seeding them for transfection.
The IS-98-ST1 strain of WNV (provided by Dr. S. Lecollinet; ANSES-ENVA, Maisons-Alfort, France) is a highly virulent strain isolated from a stork with severe neurological symptoms during the 1998 epidemic in Israel [62 (link)]. Working stocks of WNV IS-98-ST1 were generated by a single round of amplification in Vero E6 cells. Titers of virus stocks were determined by a standard serial dilution plaque assay in Vero E6 cells, and results expressed as PFU/mL [1 (link),63 (link)].
The IS-98-ST1 strain of WNV (provided by Dr. S. Lecollinet; ANSES-ENVA, Maisons-Alfort, France) is a highly virulent strain isolated from a stork with severe neurological symptoms during the 1998 epidemic in Israel [62 (link)]. Working stocks of WNV IS-98-ST1 were generated by a single round of amplification in Vero E6 cells. Titers of virus stocks were determined by a standard serial dilution plaque assay in Vero E6 cells, and results expressed as PFU/mL [1 (link),63 (link)].
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