Zebrafish IRF proteins are capable to regulate fish IFN response. (A) overexpression of zebrafish IRF proteins regulate the activation of crucian carp IFN promoter. EPC cells seeded in 48-well plates overnight were transfected with CaIFNpro-luc (100 ng), each of zebrafish IRF plasmids (100 ng) and pRL-TK (10 ng). The control cells were transfected with pEGFP-N3 (EV) instead of IRF plasmid. 30h later, cells were harvested for luciferase assays. (B) overexpression of zebrafish IRF2/4b/10 inhibits poly(I:C)-triggered activation of crucian carp IFN promoter. EPC cells seeded in 48-well plates overnight were transfected with CaIFNpro-luc (100 ng), each of zebrafish IRF2/4b/10 (100 ng), pRL-TK (10 ng) and poly(I:C) (1 ug/ml) for 30h, followed by luciferase assays. (C–E) overexpression of zebrafish IRF proteins regulate the activation of zebrafish IFNφ1 promoter (C), IFNφ3 promoter (D), and ISRE-containing promoter (E). EPC cells were transfected as in (A) with IFNφ1pro-luc (C), IFNφ3pro-luc (D), or ISRE-luc (E). (F) overexpression of zebrafish IRF2/4b/10 inhibits poly(I:C)-triggered activation of ISRE-containing promoter. EPC cells were transfected as in (B) with ISRE-luc instead of CaIFNpro-luc. The dashed line indicates the basic stimulatory effect of empty vector on IFN promoter activation. The data shown were representative of three independent experiments. (***P < 0.001, **P < 0.01).
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