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. 2017 Mar 1;18(3):528. doi: 10.3390/ijms18030528

Figure 3.

Figure 3

Figure 3

Methylation of the -1C nucleoside inhibited the binding and function of NF-κB. (a) Sequences of the oligonucleotides with tandem FABP6 or CCL2 κB sites used in DAPA. The κB site sequences were underlined, the nucleotides at -1 position were in bold; (b,c) DAPA using FABP6 (b) or CCL2 (c) oligonucleotides shown above and lysates of HEK 293T cells treated with TNFα (10 ng/mL) for 30 min. Oligonucleotides-bound NF-κB was detected by Western blot analysis using antibodies against p65 or p50, respectively. NC, negative control oligonucleotide. Input, 2% of the whole cell lysate used in DAPA; (d,e) Reporter assays of TNFα (10 ng/mL, 18 h) treated HEK 293T cells transfected with plasmids shown in the figures. Fold activations are the luciferase activities of TNFα treated group normalized to their untreated controls. The inserts shown are the relative luciferase activities of all eight reporters in each experiment. (b,c) are the results of one of three experiments. Data shown in (d,e) are means ± S.D. of the results from at least three independent experiments.