Fig. 7

Impact of calcitriol on the differentiation of MDMs from breast cancer patients (all). Expression of (A) CD14, (B) HLA-DR, (C) CD80, and (D) CD163 on M0, M1, M2a, and M2c macrophages. The cells were differentiated in the presence of M-CSF for 7 days, and the medium was replaced with fresh medium on days 3 and 6. On the eighth day, the MDMs were treated with factors differentiating them into macrophage classes: M0 (initial cells, stimulating factor: M-CSF 50 ng/mL), M1 (stimulating factor: LPS 100 ng/mL, INFγ 50 ng/mL), M2a (stimulatory factor: IL-4 50 ng/mL) and M2c (stimulatory factor: IL-10 50 ng/mL). The differentiation process was carried out in the absence or presence of calcitriol (1, 10, and 100 nM). After 48 h, the cells were treated with LPS for another 24 h. On day 11, the cells were collected for phenotypic analysis (flow cytometry analysis using the BD FACS Diva™ 6.2 program, and the fluorescence intensity was determined relative to that of the unstained control, MFI). M-CSF, macrophage colony stimulating factor; LPS, lipopolysaccharide; INFγ, interferon gamma; IL-4, interleukin 4; IL-10, interleukin 10. Additional file 1: Table S9 presents the detailed numbers of patients in each group analyzed. Statistical analysis: Dunn’s multiple comparison test; ns, nonsignificant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.