Fig. 3

LGP2-talen is not functional in fish IFN response

(A) Schematic of wild-type LGP2 (LGP2-WT), LGP2-talen, and LGP2-DH.

(B) Fish IFN promoter is activated by zebrafish LGP2 but not by LGP2-DH and LGP2-talen. EPC cells seeded in 24-well plates were cotransfected with CaIFNpro-luc (200 ng), Renilla vector (pRL-TK, 20 ng), and LGP2-WT/LGP2-DH/LGP2-talen at increasing doses (0, 10, 50, 200 ng) for 48 h, followed by luciferase assays.

(C) Cellular ifn gene was not induced by LGP2-talen. EPC cells seeded in 6-well plates were transfected with LGP2-WT, LGP2-DH, or LGP2-talen at increasing amounts (0, 40, 200, 800 ng) for 48 h, followed by RT-PCR.

(D) LGP2-talen failed to inhibit poly(I:C)-triggered IFN response. EPC cells seeded in 24-well plates were transfected for 24 h with 200 ng of CaIFNpro-luc and increasing amounts (0, 10, 50, 200 ng) of LGP2-WT or LGP2-talen or LGP2-DH, followed by transfection again with poly(I:C) (1 μg/mL). Another 24 h later, luciferase assays were performed. Western blots in (B), (C), and (D) showed the ectopically expressed LGP2 in a dose-dependent manner by zebrafish LGP2 antibody. ∗Nonspecific band. Data were shown as mean ± SD (N = 3). P values were calculated using Student’s t test. ∗∗p < 0.01. See also S3 and Table S1.