Fig. 5.

Inhibitory effect of EA/EuH on TNFα-induced MMP-9 promoter activation. a Promoter assay. MDA-MB-231 cells were transfected with 0.2 μg of promoter reporter, pMMP9-Luc(−925/+13). After 48 h, cells were treated with Ea/EuH (5 μg/mL) for 8 h, and their luciferase activities were measured. Firefly luciferase activity was normalized to Renilla luciferase activity. The data shown represent the mean ± SD (n = 9). ***, P < 0.0001. P value was analyzed by Sidak’s multiple comparisons test. b Either pMMP9-Luc(−925/+13) or pMMP9-Luc(−925/+13)mtNFκB was transfected into MDA-MB-231 cells. After 48 h, cells were treated with TNFα (10 ng/mL) for 8 h, and their luciferase activities were measured. The data shown represent the mean ± SD (n = 9). P value was analyzed by Sidak’s test. c Either pMMP9-Luc(−925/+13) or pMMP9-Luc(−925/+13)mtNFκB was co-transfected along with RelA/p65 nuclear factor kappa B (NF-κB) expression plasmid into MDA-MB-231 cells. After 48 h, cells were collected, and their luciferase activities were measured. The data shown represent the mean ± SD (n = 9). ns, not significant. ***, P < 0.0001. P value was analyzed by Sidak’s multiple comparisons test