Figure 4.

Gene silencing of Beclin-1 by pLenti-Beclin-1-shRNA (short hairpin RNA) inhibits ZIKV production. HUVEC were infected with empty lentivirus vector or pLenti-Beclin-1-shRNA. Forty-eight hours after lentivirus infection, cells were infected with ZIKV at an MOI of 1. Samples were collected at 24 h after ZIKV infection and subjected to Western blot analysis (A) or plaque assay (C). (A) Western blot analysis of Beclin-1 and viral NS3 expression. GAPDH was used as a protein loading control; (B) The ratios of NS3 to GAPDH were calculated and representative results are presented with graphs. Data are presented as means from three independent experiments. ** p < 0.01; (C) Extracellular virus yields were determined and expressed as pfu/mL. Data are presented as means ± SDs for triplicate assays. ** p < 0.01.