Functional Roles of NOD1 in Odontoblasts on Dental Pulp Innate Immunity

Figure 2

Expression and production of chemokines induced in KN-3 cells stimulated with NOD1 or NOD2 ligand. (a, b) KN-3 cells were stimulated with iE-DAP (1 or 10 μg mL⁻¹), iE-Lys (10 μg mL⁻¹), MDP (1 or 10 μg mL⁻¹), MDP-LL (10 μg mL⁻¹), or TNF-α (0.01 μg mL⁻¹) for 24 h. (a) After stimulation, total RNA was isolated and mRNA expression levels of CINC-1, CINC-2α, MCP-1, and CCL20 were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR). (b) The concentrations of CINC-1, CINC-2, MCP-1, and CCL20 in the cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA). (c) KN-3 cells were stimulated with iE-DAP (10 μg mL⁻¹), iE-Lys (10 μg mL⁻¹), or TNF-α (0.01 μg mL⁻¹) for 4, 12, or 24 h. After stimulation, total RNA was isolated and mRNA expression levels of CINC-1, CINC-2α, MCP-1, and CCL20 were analyzed by real-time RT-PCR. Values represent the means ± SDs from representative of four independent experiments and each experiment was performed in quadruplicate. Asterisks indicate significant differences ( and ) versus nonstimulated control group.

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