Figure 5.

Laboratory of genetics and physiology 2 (LGP2) interacts with melanoma differentiation-associated gene 5 (MDA5) and retinoic acid-inducible gene I (RIG-I) independent of grass carp reovirus (GCRV) infection. (A,B) LGP2 overexpression inhibits RIG-I and MDA5 promoter activities. Fathead minnow (FHM) cells were transiently transfected with 300 ng of LGP2-Flag overexpression plasmid or empty vector, 30 ng of pRL-TK, and 300 ng of report vector (RIG-Ipro-luc or MDA5pro-luc) for 16 h, and then the cells were infected with GCRV or uninfected. Dual-luciferase report assays were conducted at 24 h after GCRV infection. Error bars indicate SD (n = 4). Asterisks indicate significant differences from control (*0.01 < P < 0.05; **0.001 < P < 0.01). (C,D) LGP2 interacts with RIG-I and MDA5. FHM cells were transfected with LGP2-Flag and RIG-I-HA or LGP2-Flag and MDA5-HA for 16 h, and then infected with GCRV for 12 or 24 h. Co-IP was performed with anti-HA monoclonal antibody (Ab). Mouse IgG was used as control. WCL of each time point was subjected to IBs with anti-Flag, anti-HA, and β-Tubulin Ab, respectively.