Figure 1 | Scientific Reports

Figure 1

From: Calcitriol promotes M2 polarization of tumor-associated macrophages in 4T1 mouse mammary gland cancer via the induction of proinflammatory cytokines

Figure 1

The scheme of bone marrow-derived macrophage (BMDM) differentiation. (A) Procedure for the isolation and differentiation of BMDMs. Tumor-bearing or healthy BALB/c mice were fed diets containing various VD3 contents (normal: 1000 IU/kg, elevated: 5000 IU/kg, and deficient: 100 IU/kg) for 6 weeks. Statistically significant differences in 25(OH)D3 levels were obtained between groups before tumors were inoculated in some mice. Subsequently, calcitriol gavage (1 µg/kg) was started thrice a week for 3 weeks in mice fed with normal and deficient diets, starting from day 7 after cancer cell inoculation. All diets were continued until the day of euthanasia (in total for 10 weeks)26. (B) Generation of conditioned media from 4T1 cells treated or not with 100 nM calcitriol for 72 h in vitro. *CM from untreated (4T1 CM) and calcitriol-treated (4T1cal CM) 4T1 cells or normal culture medium were used solely during the differentiation of BMDMs from healthy mice.

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