**2. Materials and Methods**

#### *2.1. Cell Cultures and Zika Virus Strain*

Larvae lysate cells (C6/36) from mosquitos *Aedes albopictus* (ATCC CRL-1660, USA), kidney epithelial cells (Vero) from monkeys *Cercopithecus aethiops* (ATCC CCL-81), human monocytes (THP-1) from peripheral blood (ATCC TIB-202), and human endothelial cells (HMEC-1) from the dermal microvasculature (ATCC CRL-3243) were used in this study. The C6/36 cells were maintained in Leibovitz L15 medium (Biowest, Riverside, MO, USA) supplemented with 10% (*v*/*v*) fetal bovine serum (FBS; Biowest, Nuaillé, France), 10% tryptose phosphate broth (DIFCO, Lawrence, KS, USA), 1% 200 mM L-glutamine (Biowest), and 1% antibiotic solution (10,000 U/mL penicillin, 10 mg/mL streptomycin, and 25 μg/mL amphotericin B; Biological Industries, Cromwell, CT, USA) and were incubated at 28 ◦C without CO2 (Lab-Line Ambi-Hi-Low Chamber, Lab-Line Instruments Inc., Melrose Park, IL, USA). The Vero and THP-1 cells were maintained in Dulbecco's Modified Eagle medium (DMEM, Biowest) and Roswell Park Memorial Institute (RPMI) 1640 medium (Biowest), respectively. Both media were supplemented with 10% Fetal Bovine Serum (FBS), 1% 200 mM L-glutamine, and 1% antibiotic solution. The HMEC-1 cells were maintained in MCDB-131 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% FBS, 1% 200 mM L-glutamine, 10 ng/mL epidermal growth factor (Sigma-Aldrich), 1 μg/mL hydrocortisone (Sigma-Aldrich), and 1% antibiotic solution. The Vero, THP-1, and HMEC-1 cells were incubated at 37 ◦C in 5% CO2 (Series 8000 WJ CO2 Incubator, Thermo Fisher Scientific, Waltham, MA, USA). Dr. Amadou A. Sall, from Institut Pasteur Dakar, kindly provided the reference strain ZIKV MR766 (Genebank Accession HQ234498.1), with the following passage history: 146 × in suckling mouse, 1 × C6/36 cells, and 1 × Vero cells. Additionally, in our laboratory, it was passaged twice in C6/36 cells.
