FIG 2.
Inhibition of IL-1 and TLR activation of NF-κB by MC132. (A) HEK293T cells were seeded at 2 × 105 cells per ml, transfected with 80 ng NF-κB reporter gene, 40 ng TK renilla reporter gene, and 25 or 50 ng (indicated by wedge) empty vector (control) or pCEP4 plasmids expressing the indicated MCV ORFs, stimulated with 50 ng/ml IL-1β for 6 h, and then harvested and assayed for NF-κB reporter gene activity. Data are percentages of the stimulation activity for control cells and are means ± SD for triplicate samples from a representative experiment (n = 4). (B) Extracts from the samples for panel A were probed for expression of Flag-tagged viral proteins. (C to I) The same as panel A, except that instead of IL-1 stimulation, cells were transfected with 50 ng plasmid expressing TRAF6 (C), MyD88 (D), CD4-TLR9 (E and F), CD4-TLR3 (G and H), or TRIF (I) for 24 h. For panels F and H, cells were transfected with an ISRE-luciferase reporter in place of the NF-κB reporter. *, P < 0.001 compared to the control.