Figure 4.

Ouabain did not impair the entry of the virus into the cell. (A) A viral suspension containing ZIKV at multiplicity of infection (MOI) of 0.1 was incubated with equal volume of different concentrations of ouabain for 1 h at 37 °C to evaluate the virucidal activity of ouabain. Then, the suspension was titrated using the TCID₅₀ method. (B) Vero cells were infected with ZIKV (MOI = 0.1) and treated with different concentrations of ouabain for 1 h at 4C for virus adsorption. After 120 h, the supernatant was collected and titrated using the TCID₅₀ method. (C) Vero cells were incubated at 4 °C for 1 h to virus attachment (MOI = 0.1). After 1 h of adsorption, unabsorbed viruses were removed and cells were incubated at 37 °C for 1 h with different concentrations of ouabain. Then, cells were treated with 0.1 ml of citrate buffer for 1 min to inactivate adsorbed viruses but not internalized. After 120 h, the supernatant was collected and titrated using the TCID₅₀ method. Values were presented as mean ± standard deviation of three independent experiments and analyzed by one-way analysis of variance (ANOVA) followed by Tukey post-test.