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

Figure 3

Comparative effects of LDLs on marker gene expression in unpolarized RAW 264.7 M0 macrophages. M0 macrophages were treated for 24 hours in the presence of medium alone (Ctl), native LDLs (Nat), Ox-LDLs (Ox), or MpOx-LDLs (MpOx) (100 μg/mL). (a) Expression of polarization marker genes at the mRNA level (RT-qPCR). The expression of marker genes was analyzed by RT-qPCR as in Figure 1. Data are normalized with TBP used as housekeeping gene and expressed as mean fold induction relatively to Ctl cells ± SD (). (b) Expression of polarization markers at the protein level (FACS, ELISA, and WB) in Ctl and Nat-LDLs-, Ox-LDLs-, and MpOx-LDLs-treated M0 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 Ctl and treated macrophages (). GMF values are reported in arbitrary units: , 0; , 20.75; and , 33.82. The data presented are representative of 3 independent experiments. (b3) Expression of HO-1 and Srxn1 assessed by Western blotting in Ctl and Nat-LDLs-, Ox-LDLs-, and MpOx-LDLs-treated 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 .