Fig 2. TRIM25 overexpression inhibits IBDV replication.

(A-D) The influence of overexpression of pFlag-TRIM25 on IBDV replication. The DF-1 cells were transfected with 2μg pFlag-TRIM25 or pCAGGS at 24 h intervals and were infected with IBDV Gt strain at an MOI of 0.01. Subsequently, the cell and supernatant samples were collected at 24 and 48 h p.i, respectively. (A) Expression levels of TRIM25 and VP3 were determined by Western blotting. (B) Alternatively, relative intensities of VP3 were normalized to β-actin. (C) Relative fold-change of IBDV genome mRNA level of cell samples was quantified by RT-qPCR. (D) Released viral titer was detected and illustrated using a TCID₅₀ assay. All qPCR results are represented as relative fold changes after being normalized to β-actin controls. The Western blotting results are representative of one of three independently performed. Three independent experiments were performed, and data are shown as mean ± standard deviations for triplicates from representative experiments. *, P < 0.05; **, P < 0.01.