Fig. 6

Figure 6. Prophylactic Efficacy of TRIM25 Reconstitution against SVCV Infection under Sμg Conditions (A) Schematics of the experimental design. One-cell-stage embryos were microinjected with TRIM25-Myc, IFNφ1-Myc, or vector control and cultured in HEPES media for 72 h. Subsequently, embryos were challenged with about 10⁸ 50% tissue culture infectious dose (TCID₅₀)/mL SVCV at 25°C for 24 h. After washing, the embryos were subsequently cultured and monitored under Ng or Sμg conditions for six days. In addition, 30 larvae of each group were collected at 24 h post-infection (hpi), pooled, and processed for the gene expression and luciferase reporter analyses. (B) Kaplan-Meier survival curves of embryos with different treatments. Two-way ANOVA: Ng-inactivated SVCV+Vec (n = 100) versus Ng-SVCV+Vec (n = 100), p < 0.0001; Ng-SVCV+Vec (n = 100) versus Ng-TRIM25-Myc (n = 100), p < 0.0001; Ng-SVCV+Vec (n = 100) versus Ng-IFNφ1-Myc (n = 100), p = 0.0002; Ng-SVCV+TRIM25-Myc (n = 100) versus Ng-IFNφ1-Myc (n = 100), p = 0.5821; Sμg-inactivated SVCV+Vec (n = 100) versus Sμg-SVCV+Vec (n = 100), p < 0.0001; Sμg-SVCV+Vec (n = 100) versus Sμg-TRIM25-Myc (n = 100), p < 0.0001; Sμg-SVCV+Vec (n = 100) versus Sμg-IFNφ1-Myc (n = 100), p < 0.0001; and Sμg-SVCV+TRIM25-Myc (n = 100) versus Sμg-IFNφ1-Myc (n = 100), p = 0.0042. (C) qPCR analysis of SVCV glycoprotein expression. The data in the Ng-SVCV+Vec group were arbitrarily set to 1. No SVCV glycoprotein expression was detected in the Ng-inactivated SVCV+Vec group (data not shown). (D) IFNφ1 expression, IFNφ1, and NF-κB luciferase reporter assay in the indicated treatments with SVCV infection and TRIM25 or IFNφ1 rescuing. The data in the Ng-inactivated SVCV+Vec group were arbitrarily set to 1. Vec stands for empty vector. Error bars: All data points in this figure are presented as mean ± SE. Two-tailed unpaired t test. ∗p < 0.05; ∗∗p < 0.01; n.s., not significant.