Figure 4.
Figure 4. Effect of ISRE-1 site mutation on IL-12p35 gene expression. (A) RAW 264.7 cells were transiently transfected with 1 μg IL-12p35lucWT, IL-12p35lucMutA or IL-12p35lucMutB, and 80 ng pRL-TK as an internal control plasmid. Cells were stimulated with either IFN-γ (100 ng/mL) and LPS (1 μg/mL) or PolyI:C (20 μg/mL). Values represent the mean ± SEM of 6 independent experiments performed in triplicates. (B) Ectopic expression of IRF-3 5D up-regulates IL-12p35 promoter activity. RAW cells were cotransfected with 400 ng of reporter plasmid and increasing amounts of IRF-3 5D. The total amount of DNA transfected was maintained constant by complementing with the empty vector. (C) Transient transfection of IL-12p35lucWT (400 ng) in RAW cells and expression plasmids encoding IRF-1wt (400 ng) and IRF-3wt (400 ng) as indicated. Results represent the mean ± SEM of triplicates. A representative experiment of 3 is shown. RLU indicates relative light units.

Effect of ISRE-1 site mutation on IL-12p35 gene expression. (A) RAW 264.7 cells were transiently transfected with 1 μg IL-12p35lucWT, IL-12p35lucMutA or IL-12p35lucMutB, and 80 ng pRL-TK as an internal control plasmid. Cells were stimulated with either IFN-γ (100 ng/mL) and LPS (1 μg/mL) or PolyI:C (20 μg/mL). Values represent the mean ± SEM of 6 independent experiments performed in triplicates. (B) Ectopic expression of IRF-3 5D up-regulates IL-12p35 promoter activity. RAW cells were cotransfected with 400 ng of reporter plasmid and increasing amounts of IRF-3 5D. The total amount of DNA transfected was maintained constant by complementing with the empty vector. (C) Transient transfection of IL-12p35lucWT (400 ng) in RAW cells and expression plasmids encoding IRF-1wt (400 ng) and IRF-3wt (400 ng) as indicated. Results represent the mean ± SEM of triplicates. A representative experiment of 3 is shown. RLU indicates relative light units.

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