Analysis of the effect of miR-484 and miR-744 expression on DENV replication - To evaluate the effect of miR-484 and miR-744 on DENV replication, Vero cells were seeded at a density of 2 × 105 cells/well in 12-well plates. On the following day, the cells were transiently transfected with pEZX-MR03-miR-484 (Homo sapiens miR-484 stem-loop expression clone, cat. HmiR0264-MR03) or pEZX-MR03-miR-744 (H. sapiens miR-744 stem-loop expression clone, cat. HmiR0509-MR03) both obtained from GeneCopoeia, Inc. (Rockville, Maryland, USA), at a final concentration of 0.5 µg/mL using Lipofectamine 2000 according to the manufacturer’s instructions. A scrambled miRNA or empty vector (Cat. CmiR0001-MR03, Rockville, Maryland, USA) was used as the negative control. These constructs contain the GFP reporter gene facilitating the determination of transfection efficiency and the miRNA overexpression level by fluorescence microscopy. At 24 hpt, the cells were challenged with each DENV serotype separately at an MOI of 3. Alternatively, the cells were first challenged with each DENV serotype separately and after 24 h, they were transfected with pEZX-MR03-miR-484 or pEZX-MR03-miR-744 as described above. After 3 h of virus adsorption and shaking every 30 min, the medium was removed, the cells were washed twice with PBS, and the medium was replaced with DMEM supplemented with 2% FBS. The effect of either miRNA on DENV replication was assessed at 72 hpi by flow cytometry and RT-qPCR, and by western blot in the case of DENV-2. All experiments were performed in triplicate.