FIGURE 3.

Effect of pre-treatment with Spirulina extract on iNOS gene and protein expression in LPS-stimulated cortical microglia. Microglia were cultured for 24 h in 10% serum-containing medium, which was replaced with serum-free medium before pre-treatment with 100 μg/mL E1 followed by stimulation with 100 ng/ml LPS. (A) iNOS mRNA levels were quantified by real-time PCR. Data are presented as means ± SEM (n = 3 in triplicate). **p < 0.01 compared to control cells; °°p < 0.01 versus LPS stimulation. One-way ANOVA followed by Sidak’s multiple comparison test. (B) Microglia were stained with anti-iNOS antibody (red) and nuclei with DAPI (blue). Experiments were performed three times and representative immunofluorescence images are shown. Scale bar, 20 μm.