Fig 3

Fig 3 Zebrafish mylipb negatively regulates IFN-I signaling.

(A-B) Overexpression of mylipb suppressed the activity of ISRE reporter (A), zebrafish IFNφ1 reporter (Dr-IFNφ1-Luc.) (B), induced by poly I:C transfected EPC cells. We transfected EPC cells with the indicated reporters (0.1 μg/well) along with Myc-empty vector or Myc-mylipb vector (0.2 μg/well). After 24 h, we transfected the cells with poly I:C (1 μg/mL) for 24 h and then conducted luciferase reporter activity and western blot assays. (C-D) Overexpression of mylipb suppressed the activity of ISRE reporter (C), zebrafish IFNφ1 reporter (Dr-IFNφ1-Luc.) (D), induced by SVCV infection in EPC cells. We transfected EPC cells with the indicated reporters (0.1 μg/well) along with Myc-empty vector or Myc-mylipb vector (0.2 μg/well). After 24 hours, we transfected the cells with SVCV (MOI of 1) for 24 hours and then conducted luciferase reporter activity and western blot assays. (E) Overexpression of mylipb suppressed expression of ifn, isg15, and viperin induced by poly I:C in EPC cells. EPC cells were transfected with Myc-mylipb or empty vector. After 24 hours, we transfected the cells with poly I:C (1 μg/mL) for 24 hours, total RNAs were extracted for examining the mRNA levels of ifn, isg15, and viperin by qRT-PCR analysis. Western blotting tests to detect the overexpression of mylipb. (F) Overexpression of mylipb suppressed expression of ifn, isg15, and viperin induced by SVCV infection in EPC cells. EPC cells were transfected with Myc-mylipb or empty vector for 24 h and infected with SVCV (MOI of 1). After 24 h, total RNAs were extracted for examining the mRNA levels of ifn, isg15, and viperin by qRT-PCR analysis. Western blotting tests to detect the overexpression of mylipb. All data are presented as mean values based on three repeated experiments, and error bars indicate the ± SD. *,P < 0.05, **, P<0.01; ***, P < 0.001; ****, P< 0.0001.