Figure 2.

Cell viability of HBMECs infected with ZIKV. (A) HBMECs were mock-treated or cultured with ZIKV_PE243 or ZIKV_MR766, produced in C6/36 cells, in their native and inactivated forms (iZIKV). XTT metabolization was assayed at the indicated time points, and normalized according to the values obtained in cell cultures maintained in culture medium only. Data are represented as mean ± SD of six independent experiments. ^*p < 0.05 in relation to mock. (B) HBMECs were mock-treated or infected with ZIKV_PE243 or ZIKV_BR−SP and XTT assay was performed as in (A). (C,D) HBMECs were infected with ZIKV_PE243 or ZIKV_MR766. Cells were stained with AlexaFluor488-Annexin V (AnnV) and propidium iodide (PI) and were evaluated by flow cytometry, at the indicated time points. (C) Dot plot indicating the percentage of AnV⁺PI⁺ cells at each time point, in a representative experiment. (D) Average percentage of AnnV⁺PI⁺cells at each time point from three independent experiments. (E) Culture supernatants were harvested and LDH release was evaluated by measurement of NADH consumption/min in a LDH activity assay. Data are represented as mean ± SD of three independent experiments. ^*p < 0.05 in relation to mock.