Functional Roles of NOD1 in Odontoblasts on Dental Pulp Innate Immunity

Figure 4

The effects of AP-1 signal pathway in iE-DAP-stimulated KN-3 cells. (a, b) KN-3 cells were treated with PD98059 (25 μM), SB203580 (10 μM), SP600125 (10 μM), SN-50 (9 μM), or SR11302 (10 μM) for 1 h followed by stimulation with iE-DAP (10 μg mL⁻¹) for 24 h. (a) The concentrations of CINC-1, CINC-2, MCP-1, and CCL20 in cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Values represent the means ± SDs from representative of three independent experiments and each experiment was performed in triplicate. Asterisks indicate significant differences ( and ) versus noninhibitor control group. (b) mRNA expression level of CINC-2α was analyzed by real-time RT-PCR. Values represent the means ± SDs from representative of three independent experiments and each experiment was performed in quadruplicate. Asterisks indicate significant differences ( and ) versus noninhibitor control group. (c) A GFP reporter construct with transcriptional response element for AP-1 or NF-κB was transiently transfected into KN-3 cells and subjected to stimulation with iE-DAP (10 μg mL⁻¹), iE-Lys (10 μg mL⁻¹), or TNF-α (0.01 μg mL⁻¹) for 24 h. The expression level of GFP was measured using a fluorescence microplate reader (Infinite® 200 PRO, Tecan, Männedorf, Switzerland). Values represent the means ± SDs from representative of three independent experiments and each experiment was performed in quadruplicate. Asterisks indicate significant differences () versus nonstimulated control group.

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