Figure 4. Induction of β-defensin 2 triggered by TLR2, TLR4, NOD1, or NOD2 ligand in human oral epithelial cells. (a) Human oral epithelial HSC-2 cells were incubated for 8 hrs in the presence or absence of Pam₃CSSNA (100 pg/mL), lipid A (100 ng/mL), muramyldipeptide (100 µg/mL), or iE-diaminopimelic acid (100 µg/mL). After incubation, total RNA was extracted, and the mRNA expression of β-defensin 2 was analyzed by real-time PCR. (b) Human oral epithelial HSC-2 cells were incubated for 24 hrs in the presence or absence of Pam₃CSSNA (100 pg/mL), lipid A (100 ng/mL), muramyldipeptide (100 µg/mL), or iE-diaminopimelic acid (100 µg/mL). After fixation, cells were treated with anti-β-defensin 2 antiboby and then visualized with Alexa Fluor 488 (green). Nuclei were visualized by being stained with 4',6-diamino-2-phenylindole (blue). Scale bars: 20 µ m. (c) Human oral epithelial HSC-2 cells were incubated for 24 hrs in the presence or absence of Pam₃CSSNA (100 pg/mL), lipid A (100 ng/mL), muramyldipeptide (100 µg/mL), or iE-diaminopimelic acid (100 µ g/mL). The expression of β-defensin 2 was assessed by flow cytometry. Thin lines represent the isotype antibody control. The results presented are representative of 4 different experiments demonstrating similar results.