Figure 7
EMSA analysis. NF-kappaB p50 has been incubated for 15 min in the presence of increasing amounts of compounds 5 (right side of panel A), extracts from C. pyramidalis (left side of panel A), compound 21 (B-D) and reference compound 9i (right side of panel C). After the incubation of compounds (or C. pyramidalis extracts) to NF-kappaB p50, a further 15 min incubation step was carried on with a 32P-labelled double stranded oligonucleotide carrying the NF-kappaB binding sites. C. pyramidalis extracts were employed at a concentration ranging from 15 to 1000 μg/reaction as mentioned. Comparison between compound 21, compound 5, and compound 9i was performed at 0.01, 0.1, 1 an 10 mM concentrations. Concentration-dependency of the effects displayed by compound 21 was further analysed with intermediate dosages in the experiment depicted in panel D. The NF-kappaB/DNA complex were identified after polyacrylamide gel electrophoresis. Arrows indicate NF-kappaB/DNA complexes. Asterisks indicate free target DNA.