FIG. 8.

Activation analysis of transcription factors NfκB and IFN regulatory factor (IRF3). (A, B) Activation of the factors was followed 1 h postnucleofection of NIH3T3 cells with phGfΔG plasmid or after mock nucleofection. Selected confocal microscopy sections are presented. Cells were fixed and stained by (A) an antibody against nuclear factor-kappa (NF-κB) p-65 (green) and by DAPI (blue) or by (B) an IRF3-specific antibody (green) and DAPI (blue). (C) IRF3 phosphorylation in NIH3T3 cells nucleofected with phGfΔG or mock nucleofected was analyzed by Western blotting. Cells were lysed at the indicated times, separated by SDS electrophoresis, and blotted. Immunostaining was performed by phospho-IRF3 (Ser396) antibody, specific for the epitope containing phosphorylated serine at position 396. Polyclonal antibody against the IRF3 protein was used as a control.