Fig. 4
Effects of ID on the production of proinflammatory cytokines and the activation of MAPKs and NF-κB in HMC-1 cells. (a) HMC-1 cells (1 × 104 cells/well) were treated with ID (0.01 - 1 mg/ml) for 24 h and the cell viability was analyzed with MTT assays. (b-d) The production levels of TNF-α (B), IL-8 (c), and IL-6 (d) were measured using the ELISA method. HMC-1 cells (2 × 105 cells/well) were pre-treated with ID for 1 h and were stimulated with PMA (50 nM) + A23187 (1 μM) for 8 h. Values represent the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001. (e and f) Phosphorylation of p38, ERK and JNK (e) and the phosphorylation of IκBα and translocation of NF-κB (f) were assayed by western blot analysis. HMC-1 cells (1 × 106 cells/well) were incubated with various concentrations of ID and then stimulated with PMA (50 nM) + A23187 (1 μM) for 30 min (e) or 2 h (f). C, cytosol extracts; N, nuclear extracts