Fig. 3.

Screening reporter DENV against a library of 350+ ISGs. (A) Inhibition of DENV-GFP by Huh7 cells transduced with lentivirus expressing IFITM3. Data are represented by FACS plots showing uninfected (gray line) and infected (black line) cells. (B) Screening 350+ ISGs for antiviral activity against DENV-GFP. Huh7 cells were transduced with lentiviral stocks for 3 d and then infected with DENV-GFP. Cells were analyzed for ISG-mediated inhibition of DENV-GFP replication by high-throughput FACS. Data are represented as a dot plot, with replication levels normalized to a Fluc control. Selected ISGs are indicated in blue. The red line indicates the population mean. (C) Confirmation assays of select antiviral ISGs in Huh7 (Left) cells or STAT1^−/− fibroblasts (Right). Replication levels were normalized to the Fluc control. Data are presented as box and whisker plots: gray boxes extend from the 25th to the 75th percentile, with a black line at the population median; whiskers extend to show the highest and lowest values, n = 6. (D) Time course of ISG-mediated inhibition of DENV-Fluc(NS4B:L52F). ISG-expressing cells were infected with virus and Fluc levels were monitored over time. Results are presented as mean ± SD of two independent experiments, each performed in quadruplicate, n = 8 at each data point. (E–H) Individual time points of data shown in D. Statistical significance was determined by one-way ANOVA (***P < 0.001; ns, not significant).