Figure 2

zbTRIM25 potentiates RLR signaling pathway. (A,B) ZBE3 cells were transfected with pCMV-Flag-zbTRIM25 or pCMV-Flag vector for 24 h. After treated with RGNNV (A) or poly I:C (B), the transcript levels of RIG-I, MAVS, TRAF3, IRF3, IFN 1, and ISG15 mRNA in ZBE3 cells were analyzed. (C) HEK 293T cells were transfected with pCMV-Myc or pCMV-Myc-zbRIG-I together with pCMV-Flag or increasing amount of pCMV-Flag-zbTRIM25 as well as DrIFN1 pro-Luc and pRL-TK. Luciferase activities were measured and normalized to the amount of Renilla Luciferase activities. (D) ZBE3 cells were transfected with pCMV-Flag and increasing amount of pCMV-Flag-zbTRIM25 for 24 h, and then infected with RGNNV for 24 h. QRT-PCR analysis was performed for RDRP. (E,F) ZBE3 cells were either not transfected (Control) or transfected with 100 nM Control siRNA (NC) or 100 nM zbTRIM25 siRNA. Cells were then subcultured for 24 h and infected with RGNNV for 24 h. Cells were harvested subsequently. Levels of zbTRIM25 and RDRP mRNA were analyzed by qRT-PCR and normalized with 18s rRNA. Data represent the mean + SD (n = 3). Asterisks indicate significant differences between groups (*p < 0.05; **p < 0.01).