Fig. 7

Selected protein levels in calcitriol-treated CAFs derived from tumors of patients with different clinical characteristics. A–D data presented for CAFs from all patients. E representative cropped blots. F–I data presented for CAFs derived from tumors of patients with different clinical characteristics. A and F idoleamine 1 (IDO1), B and G tissue metalloproteinase inhibitor 1 (TIMP1), C and H osteopontin (OPN), D and I transforming growth factor β (TGFβ). CAFs were treated with calcitriol (cal; 1 nM or 10 nM) for 72 h. The molecular weights of the analyzed proteins were as follows: IDO1—40 kDa, OPN—40 kDa, TIMP1—28 kDa, and TGFβ1—35 kDa. Full-length blots are presented in Figure S8 in the Supplementary Materials. Patients were classified into groups according to plasma 25(OH)D₃ levels (VD3, < 30 ng/mL—deficient (n = 21), > 30 ng/mL—normal (n = 14)), plasma FSH levels (< 25.8 mIU/mL—premenopausal (n = 12), > 25.8 mIU/mL—postmenopausal (n = 23)) and regional or distant metastasis presence (if any—metastatic (n = 14), otherwise nonmetastatic (n = 21)). Densitometric analysis was performed using ImageJ software. The results were normalized to β-actin levels and untreated controls. Data are presented as the mean ± SEM. Statistical analysis was carried out using Student’s t-test or the Mann‒Whitney U test for single comparisons and one-way ANOVA or the Kruskal‒Wallis test for multiple comparisons. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001