FIG 3.

Inhibition of cytosolic DNA-sensing-pathway- and virus-stimulated NF-κB activation by MC132. HEK293T cells were seeded at 2 × 10⁵ cells per ml and transfected with the indicated reporter genes and empty vector (control) or pCEP4 plasmids expressing MC132. Cells were then infected with MVA for 16 h, and NF-κB reporter activity (A), ISRE reporter activity (B), and IFN-β promoter reporter gene activity (C) were measured. Data are percentages of the stimulation activity for control cells and are means ± SD for triplicate samples from a representative experiment (n = 4). (D and E) The same as panel A, except that cells were transfected with cGAS- and STING-expressing plasmids (25 ng each), and 24 h later, NF-κB (D) or ISRE (E) reporter gene activity was measured. (F, H, and I) The same as panel A, except that cells were transfected with 500 ng/ml poly(dA:dT) (F) or 50 ng MAVS-expressing plasmid (H) or infected with VSV for 16 h (I). (G) Extracts from the samples used for panel F were probed for expression of Flag-tagged viral proteins. (J) The same as panel A, except that cells were transfected with 10 ng, 25 ng, and 50 ng of pCEP4-MC132flag and stimulated with 10 nM PMA for 6 h. (K) The same as panel J, except that Elk1 reporter gene activity was measured. *, P < 0.001 compared to the control.