Fig 6.

Cells and C57BL/6 mice infected with C40A HSV-1 produce more IFN-β than those infected with WT HSV-1. (A and B) WT HSV-1 infection, but not C40A HSV-1 infection, affected the ubiquination of TRAF3. (A) TRAF3-Flag and HA-Ub-expressing plasmids were cotransfected into HEK293T cells, and the cells were infected with SeV, WT HSV-1, or C40A HSV-1 at an MOI of 5 for 16 h. Co-IP experiments were performed to detect the ubiquitination of TRAF3. (B) HEK293T cells were infected as in panel A, and co-IP experiments were performed using TRAF3 and Ub pAb to detect the ubiquitination of endogenous TRAF3. (C) HEK293T cells, MEF cells, and HeLa cells were infected with the indicated viruses for 16 h, and semiquantitative PCR assays were performed to detect the mRNA level of IFN-β. (D) HEK293T cells in a 24-cell plate were infected with WT HSV-1 or C40A HSV-1 at an MOI of 5 for 20 h. Medium from the cells was analyzed by ELISA for IFN-β secretion. (E) C57BL/6 mice were infected with 10⁶ PFU of the indicated virus. After 24 h, blood serum was collected and subjected to ELISA to detect IFN-β production. The data represent means ± standard deviations for three replicates. Statistical analysis was performed using the t test. *, P < 0.05.