FIG 4.
MC132 inhibits NF-κB activation by IKKs, a common convergence point in the network of signal transduction pathways activated during viral infection. (A) Schematic showing multiple signal transduction pathways to NF-κB expected to be activated during a poxviral infection, all of which were shown to be sensitive to MC132 inhibition (Fig. 1 to 3). (B and C) HEK293T cells were seeded at 2 × 105 cells per ml, transfected with 25 or 50 ng (indicated by wedge) empty vector (control) or pCEP4 plasmids expressing the indicated MCV ORFs, together with 50 ng IKKα (B) or IKKβ (C), and then harvested and assayed for NF-κB reporter gene activity 24 h later. Data are percentages of the stimulation activity for control cells and are means ± SD for triplicate samples from a representative experiment (n = 3). *, P < 0.001 compared to the control. (D) Extracts from the samples used for panel C were probed for expression of Flag-tagged viral proteins.