Fig 1: SDC1 and B-FN involvement in the VM process in ovarian cancer. (A) Flow cytofluorimetric analysis of SDC1, CD31, CA125, FBP (folate-binding protein), CD44, EpCAM (epithelial cell adhesion molecule), CD133/1, and CD117 expression in VM positive human ovarian carcinoma cell line SKOV3 and serous ovarian carcinoma cell OS13 in comparison to VM-negative clear cell carcinoma cell OS2. Gray profiles represent negative controls. In vitro Matrigel tube formation using SKOV3 ovarian cell line, OS13, and OS2 ovarian cells is reported. Scale bars, 100 µm. (B) Immunofluorescence analysis of SKOV3, OS13, and OS2 ovarian carcinoma cells stained with VE-cadherin (VE-cad), VEGFR-2, E-cadherin (E-cad), and N-cadherin (N-cad) counterstained with DAPI (4’,6’-diamidin-2-fenilindolo). Scale bars, 20 µm. (C,D) In vitro Matrigel tube formation using SKOV3 or OS13 cells in the presence of ED-B or untreated (C) and in the presence of scFv OC-46F2 or scFv anti-B-FN in comparison to untreated cells (D). The differences in tube formation for different treatments were quantified by column bar graphs. The means ± SE are indicated. Statistically differences p values between the groups connected by lines are also reported. Note: ns = no significant differences between the indicated groups.
Fig 2: Expression of SDC1 and B-FN in ovarian carcinoma. Immunofluorescence analysis of cryostat sections of SKOV3 induced in NOD SCID mice (A) and human serous ovarian carcinoma biopsy (B) for SDC1 and B-FN expression with vascular markers CD31, SMA (smooth muscle actin), and Desmin, Vascular Mimicry markers VE-cadherin (VE-cad) and VEGFR2, and ovarian cancer stem cell markers EpCAM, CD44, and CD133/1, counterstained with DAPI. The merged signals are shown. Scale bars, 20 µm.
Fig 3: Characterization of 46F2SIP (small immuno protein) antibody format. (A) Schematic representation of anti-SDC1 46F2SIP antibody format. (B) SDS-PAGE analysis of 46F2SIP in no reduction (NR) and reduction conditions (R) (left panel) and profile of elution of the antibody obtained with FPLC (Fast protein liquid chromatography) indicated that in native conditions the antibody was predominantly present in dimeric form (right panel). The molecular masses (in kilodaltons) of the standards are reported. (C) Dose-dependent curve of 46F2SIP to the immobilized human recombinant syndecan-1 extracellular domain. (D) FACS (Fluorescence-activated cell sorting) profile of 46F2SIP reactivity on SKOV3 human ovarian carcinoma cells. Gray profiles represent negative controls. (E) Immunofluorescence staining of cryostat section of SKOV3 induced in NOD SCID mice using 46F2SIP. (F) Immunofluorescence analysis on cryostat sections of SKOV3/NOD SCID for expression of SDC1, B-FN, and CD31. Nuclei were counterstained with DAPI. (G) Immunofluorescence of tumor and primary organ samples 12 h after injection of 46F2SIP and L19SIP. Antibodies were detected with anti-IgE (immunoglobulin E) antibody. While in tumors, antibodies were localized in the extracellular matrix and in vessel structures. No accumulation of antibodies was observed in organs. Nuclei were counterstained with DAPI. Scale bars, 20 µm.
Fig 4: Evaluation of SDC1 (syndecan-1) and B-FN (B-fibronectin) levels in plasma and ascites from EOC patients. (A) SDC1 plasma (pSDC1) levels in EOC patients (n = 45) were significantly higher than in healthy donors (n = 29). (B,C) Distribution of pSDC1 (plasma SDC1) levels at diagnosis in a cohort of 45 EOC patients, stratified according to stage (FIGO, International Federation of Gynecology and Obstetrics) and tumor grade, and in healthy controls. (D) SDC1 and B-FN levels in ascites from EOC patients (n = 45). Horizontal bars indicate mean ± SE (Standard error) values for each group. (E) A significant correlation (p < 0.05) was found between SDC1 and B-FN. Pearson correlation coefficient is shown (r). (F) SDC1 levels are higher in ascites than in plasma collected at the same time from a sample of 31 stage III/IV patients (p < 0.0001 by paired Student’s t-test). P values of statistically significant differences between the groups connected by lines are also reported. Note: ns = no significant differences between the indicated groups.
Supplier Page from Abcam for Human Syndecan-1 ELISA Set (without plates)