Fig. 6. Unpurified DIPs and CHT column purified DIPs can inhibit DENV-2 replication in Huh7 cells.

A Huh7 cells in triplicate wells were infected with DENV-2 at an MOI of 1 for 3 h. The virus was removed and the cells were treated with either cell-free supernatant from HEK 293T cells, HEK-DI-290 cells or DIPs from HEK-DI-290-ORF cells. For the latter two treatments, DI-290 RNA was used at five copies of DI-290 RNA/cell. Culture supernatant from each well was collected at the time points indicated (d.p.i.), and the level of DENV-2 RNA in each sample was measured by RT-qPCR using primers to detect the NS5 region. Infection in the presence of HEK 293T supernatant was set at 100%. Error bars indicate the SD. A P value comparing negative control treatments to DIPs is shown. B As in A, DENV-2 infected Huh7 cells were treated with The DIP fraction (DIP frac.), DIP in culture supernatant (DIP sup), the CHT flow-through fraction (CHT F.T. frac.) or HEK 293T supernatant (HEK 293T sup.) as a negative control. DI-290 RNA levels for the first three treatments were normalized to ten copies of DI-290 RNA/cell. At the time points indicated, the level of DENV-2 RNA in culture supernatant was measured by RT-qPCR using primers for the DENV-2 NS5 region. The P values shown were calculated with a two-tailed Student’s t test. A representative of three experiments with similar results is shown.