Generation and characterization of murine bone marrow derived M0, M1 and M2 macrophages.

(A) Scheme for the generation of mouse bone marrow derived macrophages (BMDMs, M0) and subsequent polarization of M1 (100 ng/ml LPS) and M2 (20 ng/ml mIL-4) macrophages. (B) Fluorescent images show M0, M1 and M2 macrophages containing latex beads (red). Actins were stained with FITC-phalloidin (green). Nuclei were counterstained with Hoechst (blue). Note the significantly increased phagocytosis capacity of M1 macrophages. Bar, 20 μm. (C) Quantification of phagocytosis. Data was calculated as bead per cell from 5 randomly chosen fields. ** p < 0.01 by one-way ANOVA with post-hoc Tukey HSD Test. (D) The target gene expression profiles of naïve and differentially polarized macrophages. Fold expression is calculated relative to the internal control of GAPDH mRNA expression. (E) Differential M1 (IL-1β and iNOS) or M2 marker (IL-10 and TGFβ) gene expressions induced by LPS or mIL-4, respectively. Fold change is calculated as M2/M1 ratio of mRNA expression.

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