Figure 6

In vitro analyses of the YFV NS5 (ID)- eIF3L interaction. (A) pGEX 5X1-ID plasmid, the GST-fused ID (33 kDa) expressed in E. coli BL21 (DE3). (B) Control GST (26 kDa) expressed in E. coli BL21 (DE3). (C) HeLa cells were transfected with a plasmid expressing Flag-tagged eIF3L (67 kDa) (column 1). This protein was incubated with resin-bound GST or GST-ID. After the samples were washed, the resin-bound proteins were resolved by SDS-PAGE and then analyzed by a western blot assay using an anti-Flag antibody. The interaction of Flag-eIF3L with GST-ID (column 3) was confirmed, without a nonspecific interaction with GST (column 2).