Fig. 2

Tumor tissue and TAM characteristics—selected protein expression. (A) EpCAM expression in tumor tissue from patients with various 25(OH)D₃ plasma levels. N = 23 normal; N = 57 deficient. (B) Tumor tissue expression of transforming growth factor β (TGFβ; N = 32), epithelial cell adhesion molecule antigen (EpCAM; N = 39–40), osteopontin (OPN; N = 39–40) and (C) IRF4 (N = 38–40) in patients with low and high CYP27B1 (1α-hydroxylase) expression in tumor tissue. (A-C) Proteins expression was analyzed by western blot and protein tested/β-actin ratio is provided. (D) Representative blots showing EpCAM expression. (E) Representative blots showing TGFβ, OPN, and IRF4 expression. The patients were described as (D) for plasma 25(OH)D₃ concentration (normal or deficient) and (D) and (E) for CYP27B1 expression (high + and low −). TAMs expression of (F) HLA-DR and CD44 isolated from metastatic and nonmetastatic patients (N = 14–36) and of G CD11b, CD80, HLA-DR, CD163, CD200R, CD204, and CD44 in patients with low and high CYP27B1 expression in tumor tissue (N = 21) were evaluated. (H) Gating strategy for flow cytometry analyses; CD11b staining is shown as an example. (F-H) The data were visualized using Flowing Software version 2.5.1. (https://flowingsoftware.com) and the fluorescence intensity was determined relative to that of the unstained control (MFI). Statistical analysis: Mann‒Whitney test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.