Myeloperoxidase-Oxidized LDLs Enhance an Anti-Inflammatory M2 and Antioxidant Phenotype in Murine Macrophages

Figure 1

Characterization of RAW 264.7 M0, M1 (LPS + IFN-γ), and M2 (IL-4 + IL-13) polarized macrophages. Cells were polarized as described in Materials and Methods. (a) Expression of polarization marker genes at the mRNA level (RT-qPCR), in M0, M1, and M2 macrophages. The expression of iNOS and IL-6 as M1 markers, Arg1 and MRC1 as M2 markers, and HO-1 and Srxn1 as MOX redox-sensitive markers was analyzed by RT-qPCR. Data were normalized with TBP used as housekeeping gene and expressed as mean fold induction relatively to M0 cells ± SD (). (b) Expression of polarization markers at the protein level in M0, M1, and M2 macrophages. (b1) Production of secreted IL-6 (M1 marker) in cell culture supernatants assessed by ELISA. Data are expressed relatively per μg of protein per well as mean ± SD (). (b2) Surface expression of MRC1 (M2 marker) analyzed by flow cytometry in M0, M1, and M2 macrophages (). GMF (geometric mean of the fluorescence intensity) values are reported in arbitrary units: , 65.57 and , 28.9. The data presented are representative of 3 independent experiments. (b3) Expression of HO-1 and Srxn1 assessed by Western blotting in M0, M1, and M2 macrophages. Data are normalized with TBP used as loading control and expressed as mean ± SD (). Results are representative of 3 independent experiments. ANOVA 1: and .