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. 2017 Mar 23;12(3):e0174441. doi: 10.1371/journal.pone.0174441

Fig 6. Effects of Nω-Nitro-L-Arginine (NNA) on IL-1β inducible NF-κB promoter activity, expression and NO production.

Fig 6

Caco-2 cells transfected with pNF-κB-Luc vector were treated with IL-1β (4 ng/ml) for 4 hours after pretreatment of NNA (0.5 mM) and L-Arg (5 mM) for 4 hours (Fresh medium with NNA and L-Arg was applied after removing medium for pre-treatment). Cells were harvested for luciferase activity and isolations of protein and total RNA. A: NF-κB luciferase activity is expressed as fold-induction of normalized luciferase activity. B: NF-κB mRNA were measured by qRT-PCR, then normalized to β-actin. C: NF-κB p65 protein was measured by Western blot analysis normalized to β-actin. D: NO levels in cell culture media were measured as described in Methods. NNA alone did not affect the LPS-induced NF-κB promoter activity, NF-κB mRNA and protein expression levels (not shown). Data are means ± SE, * P<0.05. (n = 4–6 /group).