FIG 2.
UL94 negatively regulates viral DNA-triggered signaling. (A) UL94 inhibits HCMV, HSV-1, or VACV-triggered transcription of antiviral genes in HFFs. HFFs stably expressing UL94 (4 × 105) were left uninfected or infected with HCMV (multiplicity of infection [MOI] = 1), HSV-1 (MOI = 1), or VACV (MOI = 1) for 6 h before qPCR analysis. (B) Effects of UL94 on transcription of antiviral genes induced by transfected dsDNA in HFFs. HFFs stably expressing UL94 (4 × 105) were transfected with the synthesized dsDNA representing a 60-bp or 120-bp fragment of HSV-1 genome (HSV60 or HSV120, respectively) or a 70-bp fragment of VACV genome (VACV70) (1 μg/ml) for 6 h before qPCR analysis. (C) Effects of UL94 on SeV-induced transcription of antiviral genes in HFFs. HFFs stably expressing UL94 (4 × 105) were left uninfected or infected with SeV (MOI = 1) for the indicated times before qPCR analysis. (D) Effects of UL94 on secreted IFN-β in HFFs. HFFs stably expressing UL94 (4 × 105) were left uninfected or infected with HCMV (MOI = 1), HSV-1 (MOI = 1), or SeV (MOI = 1) for 12 h before measurement of IFN-β by ELISA. (E) Effects of UL94 on the phosphorylation of TBK1 and IRF3 induced by HCMV and transfected HSV120 in HFFs. HFFs stably expressing UL94 (4 × 105) were left uninfected or infected with HCMV (MOI = 1), or transfected with HSV120 (1 μg/ml) for the indicated times before immunoblot analyses were performed with the indicated antibodies. (F) Effects of UL94 on SeV-induced phosphorylation of TBK1 and IRF3 in HFFs. HFFs stably expressing UL94 (4 × 105) were left uninfected or infected with SeV (MOI = 1) for the indicated times before immunoblot analyses were performed with the indicated antibodies. Graphs show means ± SDs, n = 3. *, P < 0.05; **, P < 0.01 (unpaired t test).