Figure 4.

The replication of duck Tembusu virus (DTMUV) is limited by TRIM25. (A) Expression of TRIM25 by immunofluorescence. Duck embryo fibroblasts (DEFs) were transiently transfected with the pcDNA3.1-c-myc-TRIM25 plasmid. Nuclei were stained with DAPI. (B) Cell viability assays after transfection with TRIM25 or empty vector. (C–F) TRIM25 ectopic expression inhibited DTMUV replication. The expression of viral RNA (C) and proteins (D) was analyzed in DTMUV-infected DEFs with or without TRIM25 overexpression. (E) Titers of DTMUV in the supernatants of infected DEFs with or without TRIM25 transfection were measuredusingTCID₅₀ assays. (F) Immunofluorescence analysis of DTMUV proteins in DEFs. DEFs were transfected with TRIM25 or an empty vector for 24 hand infected with DTMUV for 24 h. Immunofluorescence was performed using E/NS5 mouse polyclonal antibody and c-myc mouse antibody.