Fig. 5
Inhibition of NF-κB activity is critical for β-Asarone induced proliferation inhibition and apoptosis. a Raji cells were treated with 100 μM β-Asarone for 72 h. Cells were subjected to nuclear protein or total protein extraction, then western blot analysis was performed. b The treatment was the same as a. Cells were collected and perform the luciferase reporter assay. c, d Raji cells were treated with or without 100 μM β-Asarone and (or) 10 ng/ml TNF-α for 72 h. Cells were subjected to nuclear protein extraction and western blot analysis (c) or luciferase reporter assay (d) (*p < 0.05, **p < 0.01, ***p < 0.001, the ANOVA test, followed by Least Significant Difference test, were used to make statistical comparisons). e, f Raji cells were expressed IκBα shRNA via lentivirus-mediated gene knockdown. Cells were collected for western blot analysis (e) or luciferase reporter assay (f). g, h Raji cells were expressed IκBα shRNA via lentivirus-mediated gene knockdown. Cells were treated with or without the indicated concentration of β-Asarone. Cells were collected for western blot analysis (g) or luciferase reporter assay (h). i Raji cells expressed IκBα shRNA via lentivirus-mediated gene knockdown. IκBα shRNA expressed cells and control cells were treated with or without the indicated concentration of β-Asarone. Cell counting assay was performed at the indicated time points. Bars represent mean ± SD of three independent experiments. j Apoptosis was evaluated by the Annexin V-FITC/PI staining and flow cytometry analysis. Representative results were shown in the left panel and statistical results were shown in the right panel. The bar represents mean ± SD of three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001, the ANOVA test, followed by Least Significant Difference test, were used to make statistical comparisons). k Raji cells were expressed IκBα shRNA via lentivirus-mediated gene knockdown. IκBα shRNA expressed cells and control cells were treated with or without the indicated concentration of β-Asarone. ALDEFLUOR assay was performed at the indicated time points. Bars represent mean ± SD of three independent experiments