FIG 2.

Screening of DEV viral proteins that modulate the cGAS-STING pathway. (A) DEFs were cotransfected with the IFN-β Luc reporter, pRL-TK, and the empty vector or cGAS-HA and STING-HA combined. The luciferase activity was measured at 36 h posttransfection. (B) DEFs were cotransfected with the IFN-β Luc reporter, cGAS-HA, and STING-HA expression plasmids and then inoculated with DEV (MOI = 0.01). The luciferase activity was measured at 12 h and 24 h postinfection. (C) DEFs were cotransfected with cGAS-HA and STING-HA expression plasmids and then inoculated with DEV (MOI = 0.01). The IFN-β mRNA level was measured by real-time qPCR at 12 h and 24 h postinfection. (D) DEFs were transfected with the IFN-β Luc reporter and pRL-TK, together with cGAS-HA, STING-HA, and an DEV ORF expression plasmid or the empty vector. The IFN-β luciferase activity was measured at 36 h posttransfection. The data of DEV ORFs and the empty vector from three independent tests are presented as a heat map. Higher IFN-β Luc activation levels are indicated by red, whereas lower levels are indicated by green, which corresponds to a higher degree of inhibition. (E) Various doses of the top five DEV viral protein inhibitors and the empty vector were cotransfected with cGAS-HA and STING-HA expression plasmids into DEFs, and IFN-β Luc activity was measured at 36 h posttransfection. (F) The top five DEV viral protein inhibitors and the empty vector were cotransfected with cGAS-HA and STING-HA expression plasmids into DEFs. The IFN-β mRNA levels were measured by real-time qPCR at 36 h posttransfection. The fold changes were compared with those of the empty vector controls. All controls and treated groups were performed and examined in triplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001.