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. 2019 Apr 16;23(6):4301–4312. doi: 10.1111/jcmm.14322

Figure 2.

Figure 2

Xanthatin blocked NF‐κB signalling by directly inhibiting IKKβ kinase activity. A, Effects of xanthatin on the TNFα‐induced luciferase activity. HEK293/NF‐κB cells were pre‐treated with xanthatin at indicated concentrations for 1 h and luciferase activity was measured following stimulation of TNFα (1 ng/mL) for 4 h. B, Xanthatin dose‐dependently inhibited TNFα‐induced NF‐κB pathway. MDA‐MB‐231 cells were pre‐treated with xanthatin for 1 h before stimulation by TNFα (1 ng/mL) for 10 min. Cell lysates were processed for Western blot analysis. C, Xanthatin time‐dependently inhibited TNFα‐induced NF‐κB pathway. MDA‐MB‐231 cells were pre‐treated with 20 μmol/L xanthatin for various durations (0‐120 min) before stimulation by TNFα (1 ng/mL) for 10 min. Cell lysates were processed for Western blot analysis. D, IKKβ in vitro kinase assay. The overexpressed IKKβ protein immunoprecipitated from the HEK293 was subjected to in vitro kinase assay as described in Materials and Methods. (n = 3, *P < 0.05, **P < 0.01, ***P < 0.001 compared with TNFα‐induced cells)