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Fig. 5 | Molecular Medicine

Fig. 5

From: Characterization and Functional Analysis of the Human Inducible Nitric Oxide Synthase Gene Promoter

Fig. 5

Comparison of the luciferase activity using the human 1800-bp promoter and nitric oxide production of A549 cells activated with various cytokines alone or in combination

Equal numbers of A549 cells were transfected with the human 1800-bp reporter construct followed by cytokine activation. Aliquots of the transfected cells were collected 8 hr after activation and analyzed for both luciferase activity (open bars) and NO production by the arginine to citrulline assay (solid bars). Cytokines were added singly or in combination using 100 µ/ml IL-1β, 10 ng/ml of TNFα, or 500 µ/ml of IFNγ. Luciferase activity is presented as the percentage increase over that seen in nonactivated cells. Nitric oxide production is presented as picomole of [3H]-citrulline produced per milligram of cellular protein. Culture supernatant concentrations of nitrite were determined by the Griess reaction and found to correlate with that determined by the arginine to citrulline assay (data not shown).

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