Fig. 3.

iNOS regulation of IRF1 transcriptional activity. a 293 T cells were transfected with plasmids pcDNA3 or pcDNA3-hiNOS for 24 h, and treated with L-NIL (100 μM, 24 h). The iNOS/NO-induced nuclear IRF1 was analyzed by Western blot (upper). Similar as (upper), but the NO production was detected (lower). b Similar to (a) upper, but the iNOS/NO induced-IRF1 was evaluated by immunofluorescence staining, green: iNOS or IRF1. c Human hepatocytes were infected by AdhiNOS or Adlacz for 24 h. Representative images of immunofluorescence staining are shown for IRF1 expression, green: iNOS; red: IRF1. d 293 T cells were transfected with pT109-IRF1 (0.5 μg) and iNOS expression plasmid, pT109-IRF1-iNOS with different concentrations of iNOS. Total amounts of plasmid DNA were kept constant by adding the empty pcDNA3A vector. Transcriptional activities of IRF1 were analyzed by luciferase assay (RLA: relative luciferase activity), cells transfected with pT109-IRF1-iNOS vs. pT109-IRF1, *P = 0.0003. The data shown are representative of three experiments with similar results