Journal: BMC Cancer

Article Title: Dual effect of vitamin D 3 on breast cancer-associated fibroblasts

doi: 10.1186/s12885-024-11961-z

Figure Lengend Snippet: 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 S in the Supplementary Materials. Patients were classified into groups according to plasma 25(OH)D 3 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

Article Snippet: The following primary antibodies (with respective dilutions) were used: rabbit anti-idoleamine 1 (IDO1) polyclonal antibody (1:1000), rabbit anti-OPN polyclonal antibody (1:1000), rabbit anti-TGFβ1 polyclonal antibody (1:500) and rabbit anti-TIMP1 polyclonal antibody (1:1000, all from Proteintech, Rosemont, IL, USA).

Techniques: Derivative Assay, Clinical Proteomics, Software

Journal: BMC Cancer

Article Title: Dual effect of vitamin D 3 on breast cancer-associated fibroblasts

doi: 10.1186/s12885-024-11961-z

Figure Lengend Snippet: Selected mRNA expression in calcitriol-treated CAFs derived from tumors of patients with different clinical characteristics. CAFs were treated with calcitriol (cal; 1 nM or 10 nM) for 72 h. A1 – G1 data presented for CAFs from all patients. A2 – G2 data presented for CAFs derived from tumors of patients with different clinical characteristics. A – C —C motif chemokine ligand 2 (CCL2), B metalloproteinase 9 (MMP9), podoplanin (PDPN), secreted phosphoprotein 1 (SPP1), tissue metalloproteinases inhibitor 1 (TIMP1), tenascin C (TNC), vitamin D receptor (VDR). Patients were classified into groups according to plasma 25(OH)D 3 levels (VD3, < 30 ng/mL—deficient ( n = 29), > 30 ng/mL—normal ( n = 15)), plasma FSH levels (< 25.8 mIU/mL—premenopausal ( n = 15), > 25.8 mIU/mL—postmenopausal ( n = 29)) and regional or distant metastasis presence (if any—metastatic ( n = 15), otherwise nonmetastatic ( n = 29)). mRNA expression was calculated according to the ΔΔCt comparative method and normalized to the untreated control. 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

Article Snippet: The following primary antibodies (with respective dilutions) were used: rabbit anti-idoleamine 1 (IDO1) polyclonal antibody (1:1000), rabbit anti-OPN polyclonal antibody (1:1000), rabbit anti-TGFβ1 polyclonal antibody (1:500) and rabbit anti-TIMP1 polyclonal antibody (1:1000, all from Proteintech, Rosemont, IL, USA).

Techniques: Expressing, Derivative Assay, Clinical Proteomics, Control

Journal: BMC Cancer

Article Title: Dual effect of vitamin D 3 on breast cancer-associated fibroblasts

doi: 10.1186/s12885-024-11961-z

Figure Lengend Snippet: 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 S in the Supplementary Materials. Patients were classified into groups according to plasma 25(OH)D 3 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

Article Snippet: The following primary antibodies (with respective dilutions) were used: rabbit anti-idoleamine 1 (IDO1) polyclonal antibody (1:1000), rabbit anti-OPN polyclonal antibody (1:1000), rabbit anti-TGFβ1 polyclonal antibody (1:500) and rabbit anti-TIMP1 polyclonal antibody (1:1000, all from Proteintech, Rosemont, IL, USA).

Techniques: Derivative Assay, Clinical Proteomics, Software