Fig. 3. DVG-A inhibits WT virus production in vertebrate and invertebrate cells.

a Schematic diagram of the engineered DVGs. For DVG-A and DVG-A out-of-frame, the respective nucleotide and amino acid sequences are shown. Symbol (*) refers to a stop codon. The deleted nucleotide in DVG-A out-of-frame is crossed in red. For DVG-B and DVG-C, the nucleotides at the deletion junction are indicated. b Virus titers 72 h following co-transfection of DVG and WT-encoding plasmids in HEK-293T cells at different DVG:WT molar ratios (1:10, 1:1, and 10:1) are shown. More WT-encoding plasmid or control DNA were used as controls. ****p ≤ 0.0001 (compared to WT + WT conditions by two-way ANOVA with Dunnet’s multiple comparison). c Vero cells (commonly used for replicon assays) were transfected with WT replicon RNA complemented with control RNA, inactive replicon, DVG-A or DVG-A out-of-frame. Replicon activity was measured at the indicated time points p.t. ****p ≤ 0.0001 (compared to RLU values in WT replicon only-transfected cells, by two-way ANOVA with Dunnet’s multiple comparison). d Inhibition assays in mosquito cells. C6/36 (left) or U4.4 (right) cells were transfected with the indicated amount of control RNA, DVG-A or DVG-A out-of-frame. 18 h later, cells were infected with a MOI of 0.1 PFU/cell and virus titers measured 5 days p.i. ****p ≤ 0.0001 (compared to untransfected cells by two-way ANOVA with Dunnet’s multiple comparison). e Inhibition assays in cells knocked down for Ago-2 expression. U4.4 cells were transfected with Ago-2 or control dsRNA and the indicated RNA. 18 h later, cells were infected at an MOI of 0.1 PFU/cell and virus titers measured at 5 days p.i. ****p ≤ 0.0001 (compared to control RNA-transfected cells by two-way ANOVA with Dunnet’s multiple comparison). All graphs show the mean and SD; n = 3 per group for a representative experiment of three. Control RNA: pTRI-Xef RNA; control DNA: GFP-expressing plasmid. Source data are provided as a Source Data file.