Immunophenotype of MOG-immunized M1 and M2 macrophages obtained from CFA mice. Monocytes obtained from spleen of CFA mice were differentiated into macrophages using M-CSF (50 ng/mL) for 6 days and then polarized into M1 with IFN-γ (10 ng/mL) plus LPS (1 μg/mL) or into M2 with IL-4 (20 ng/mL) for 2 more days in the presence of myelin oligodendrocyte glycoprotein epitope (MOG35-55). (A) Expression of markers by flow cytometry upon staining cells at cell surface (CD40 and CD86) or intracellularly (iNOS). Data are reported as percentage of positive cells and are representative of four independent experiments ± SEM. * p < 0.05, ** p < 0.01 (B) ELISA of cytokines and chemokines (IL-6, IL-12, TNF-α and CCL2) in supernatants of MOG-immunized M1 macrophages obtained from CFA mice. Data are reported as pg/mL and represent eight independent experiments means ± SEM. * p < 0.05 (C) Expression of markers by flow cytometry upon staining cells at cell surface (CD40, CD80 and CD86). Data are reported as percentage of positive cells and are representative of eight independent experiments ± sem. * p < 0.05, (D) ELISA of chemokines (CCL2, CCL3, CCL4, and CCL22) in supernatants of M2 macrophages. Data are reported as pg/mL and represent eight independent experiments means ± SEM. * p < 0.05, *** p < 0.001.