Fig 8.

YB-1 knockdown decreases the association of NS3, DDX3, DDX6, and IGF2BP2 with LDs. Huh7.5 cells transduced with shRNA-expressing lentiviruses were transfected with a JFH-1-expressing plasmid. (A) Three days later, cells were collected and LDs were purified with a sucrose-based flotation assay. The LD fractions were analyzed by Western blotting with anti-ADRP, anti-core, anti-NS3, anti-DDX3, anti-DDX6, and anti-IGF2BP2 antibodies. (B) Cells were treated as in panel A, fixed, and probed with anti-NS3 and anti-core antibodies. Nuclei and LDs were stained with Hoechst dye and LipidTOX, respectively. Cells were analyzed by laser scanning microscopy. (C) Percentages of colocalization of NS3 or core with LDs were calculated for each condition with MetaMorph image analysis software and plotted.