Fig 4.

HCV induces the relocalization of DDX3, DDX6, IGF2BP2, LARP1, and C1QBP to core-containing LDs. (A to G) JFH-1-expressing Huh7.5 cells were fixed and probed with antibodies that recognize the indicated host and viral proteins. Nuclei and LDs were stained with Hoechst dye and LipidTOX, respectively. Unmerged and merged higher-magnification images of the boxed areas are shown. Yellow and white arrows indicate JFH-1-expressing cells and uninfected cells, respectively. (H) Huh7.5 cells were transfected with JFH-1 and the control vector. Five days later, cells were treated with 200 μM BSA-complexed oleic acid. Sixteen hours later, cells were collected and LDs were purified with a sucrose-based flotation assay. Aliquots of lysed cells and the LD fraction were analyzed by Western blotting with anti-ADRP, anti-core, anti-NS3, anti-YB-1, anti-DDX3, anti-DDX6, anti-IGF2BP2, anti-C1QBP, anti-hnRNG-T, and anti-actin antibodies.