FIGURE 5.

Cholesterol is required for Zika virus (ZIKV) infection. (A,D) Cell viability upon methyl-β-cyclodextrin (MβCD) and filipin treatment was assessed, as described in the text. (B,E) Treatment with MβCD or filipin inhibited ZIKV infection. T98G cells were pretreated with increasing concentrations of MβCD or filipin for 1 h at 37°C. The cells were extensively washed, followed by infection with ZIKV at a multiplicity of infection (MOI) of 5. At 24 h post-infection, the infected cells were lysed to quantitate viral RNA copy numbers by RT-qPCR. (C,F) At 48 h post-infection, the titers of supernatant viruses were determined by plaque assay. (G) The cells were pretreated with 7.5 mM MβCD for 1 h, extensively washed, and then infected with ZIKV at an MOI of 5. At 24 h post-infection, the cells were fixed and stained with an anti-ZIKV envelop antibody (green). The cells were fixed and stained with DAPI (blue). (H) The cells were pretreated with 1 μm filipin for 1 h, extensively washed, and then infected with ZIKV at an MOI of 5. At 24 h post-infection, the cells were fixed and stained with an anti-ZIKV E antibody (green). Nuclei were counterstained with DAPI (blue). Scale bars in all panels represent 50 μm. Representative confocal images from three independent experiments are shown (G,H). The data shown are the mean ± SD of three independent experiments (A–F). *P < 0.05; **P < 0.01; ***P < 0.001.