Additional file 1 of Stromal cartilage oligomeric matrix protein as a tumorigenic driver in ovarian cancer via Notch3 signaling and epithelial-to-mesenchymal transition
Creators: name: Gorji-Bahri, Gilar
name: Krishna, B. Madhu
name: Hagerling, Catharina
name: Orimo, Akira
name: Jirström, Karin
name: Papadakos, Konstantinos S.
name: Blom, Anna M.
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Mathematical Sciences not elsewhere classified
subject: Developmental Biology
subject: Cancer

description: Additional file 1: Figure S1. A and BKaplan-Meier plots, obtained from the online KM plotter, representing the correlations between the higher expression of COMP with a shorter OS and progression-free survival of serous ovarian cancer patients. C Levels of COMP expression by stromal cells in high-grade serous tubo-ovarian cancer, extracted from ScPanStroma database. D The positive correlation between COMP and different isoforms of TGFB (B1, B2, and B3) in Ovarian Serous Cystadenocarcinoma, extracted from cBioPortal database. E The positive correlation between COMP and different isoforms of TGFB (B1, B2, and B3) in ovarian tumor dataset, extracted from GEPIA2 database using spearman correlation coefficient. OS: Overall survival, PFS: Progression-free survival. Figure S2. Graphical schemes representing the co-culture assay of COMP-expressing CAFs with ovarian cancer cell lines A and CAFs treatment with TGF-β isoforms B. C, D Cell proliferation assay for SKOV3 and OAW42 cells treated with increasing concentrations of COMP using CyQUANT cell proliferation assay kit. Data were normalized to PBS-treated cells. The p-value was calculated by one-way ANOVA followed by Dunnett’s post-test (n=3, mean±SD). E. Cell proliferation assay for CFSE-labeled SKOV3 cells cocultured with COMP-expressing CAFs or the mock control CAFs using flowcytometry. The p-value was calculated by two-way ANOVA followed by Sidak’s post-test (n=3, mean±SD). F Apoptosis assay using Annexin V-APC and Zombie aqua dye in four samples, including BSA (20 μg/ml)-treated, COMP (20 μg/ml)-treated, cisplatin-treated, and COMP in combination with cisplatin-treated (COMP+cisplatin) of SKOV3 and OAW42 cells. SKOV3 and OAW42 cells were treated with 10 μM and 20 μM of cisplatin, respectively. The p-value was calculated by one-way ANOVA followed by Sidak’s post-test (n=3, mean±SD). G Evaluation of total β-catenin and pGSK3β protein expression levels in SKOV3 cells treated with COMP (20 and 50 μg/ml) or BSA (50 μg/ml) as a control by western blotting. PBS-treated cells were also used as a control. β-tubulin served as a loading control. The p-value was calculated by one-way ANOVA followed by Dunnett’s post-test. The data are representative of three independent experiments and graphs depict mean±SD. (*, **, and ns indicate p&lt;0.05, p&lt;0.01, and non-significant, respectively). Table S1. List of the used antibodies. Table S2. Significant deregulated EMT-specific genes in SKOV3 cell line treated with recombinant COMP (20 μg/ml) or BSA (20 μg/ml) obtained by RT-qPCR array. Table S3: Significant deregulated EMT-specific genes in OAW42 cell line treated with recombinant COMP (20 μg/ml) or BSA (20 μg/ml) obtained by RT-qPCR array.