Figure 5

The TBK1 binding motif of STING, not IRF3, determines the DP71L-STING interaction. A PK-15 cells were transfected with DP71L-Flag plasmids in dose-dependent manner. After 24 h, cells were stimulated with 4 µg/mL of cGAMP for an additional 12 h. Subsequently, the WCLs were subjected to STING IP and bound proteins were detected by immunoblotting with indicated antibodies. B DP71L-Flag and control vector stably expressing 3D4/21 cells were infected with HSV-GFP (MOI = 1.0) and harvested at the indicated time points. WCLs were subjected to IP with endogenous STING and associated protein levels were detected by immunoblotting. C HEK293T cells were transfected with control, DP71L-Flag, and Strep-tagged STING mutant plasmids (S366A, P371Q, L374A, and R375A). Strep PD assay was performed, followed by immunoblotting with indicated antibodies. D Graphical summary of DP71L immune evasion. ASFV DP71L evades antiviral immune responses targeting the central immune molecule STING. Data are representative of at least two independent experiments, each with similar results. Protein sizes are expressed in kDa. The values are expressed as means ± SD for two biological replicates. Student’s t-test: *, P < 0.05; **, P < 0.01; ns, not significant.