1. From: Effect of Mutant IκB on Cytokine-Induced Activation of NF-κB in Cultured Human RPE Cells
Invest. Ophthalmol. Vis. Sci ;44(3): doi: /iovs
Figure Legend:
Inhibition of NF-κB transcriptional activity in RPE cells infected to express the mutant IκB. RPE cells in triplicate wells were cotransfected with 10 μg plasmid DNA encoding firefly luciferase driven by the NF-κB promoter and 2 μg of DNA encoding a constitutively expressed renilla luciferase. The firefly luciferase plasmid contains a luciferase reporter gene driven by a tandem repeat of NF-κB binding elements and therefore serves as an experimental reporter of NF-κB activation. Renilla luciferase serves as control reporter to normalize the activity of the experimental reporter, based on transfection efficiency and cell viability. Two days after transfection, the cells were infected with adenovirus containing LacZ (control virus, MOI = 10) or mutant IκB (MOI = 10). Noninfected cells were included as an additional control. One day after infection, cells were exposed to medium alone or IL-1β (5 U/mL) for 4 hours. Cells were harvested, and lysates were tested for luciferase activity with a dual-luciferase assay system. Relative luminescence reflects the ratio of firefly luciferase activity (NF-κB-dependent) to renilla luciferase (control) activity to standardize for transfection efficiency. Results are expressed as the mean ± SD of results in three experiments. *P < 0.01 versus untreated cells; **P < 0.01 versus IL-1β-treated cells in no virus group or control virus group; #P < 0.01 versus untreated cells in control virus group.
Date of download: 10/22/2017
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