Figure 2.

Interferons potentiate NFκB activation in response to the viral PAMP poly(I:C). (A) Electrophoretic mobility shift assay (EMSA) of nuclear NFκB activity in wild-type and ifnar^−/− BMDMs stimulated with LPS and poly(I:C). Quantitated activity is indicated below each band. (B) EMSA of nuclear NFκB activity in TEPMs cultured with or without IFNγ for 24 h prior to exposure to poly(I:C) or LPS. (A,B) show data representative of three biological replicates. Quantitations of phosphorimager data are relative to peak activity in controls which is set to 1. (C) Single-cell tracking of RelA-mVenus localization in 577 Poly(I:C) stimulated BMDMs cultured in the absence or presence (24 h) of IFNβ and IFNγ. Nuclear NFκB activity is indicated as nuclear:cytoplasmic ratio. The time-course response of each tracked cell is displayed as a row in the heatmap with brighter colors corresponding with increasing nuclear localization of NFκB. (D) The average nuclear NFκB activity of 577 tracked cells is shown for naïve and IFNβ- and IFNγ-primed conditions. (C,D) show data representative of two biological replicates.