Fig. 2: OvCa cells induce Jagged2 expression early in metastasis to the omentum.

A Schematic diagram of SKOV3-ip cell injection in mouse peritoneum. B Locations of (i) ascites, (ii) omentum, (iii) primary ovary, and (iv) diaphragm tumor progression 3- weeks after i.p. injection (left panel). H & E staining of omentum without (top right panel) and with tumor (right bottom panel). C Endpoint analysis of malignant ascites and total number of tumor nodules in omentectomized mice 4 weeks after intraperitoneal cell injection of SKOV3-ip cells. Data are shown as Mean +/-SEM (n = 3). Statistical significance was calculated using Kruskal-Wallis one-way ANOVA followed by Dunn’s multiple comparison test. **P < 0.01, ****P < 0.0001. D–F Jagged2 protein and RNA expression in whole mouse omentum three weeks following in vivo i.p. injection of SKOV3-ip tumor cells. Immunofluorescence (D), Immunoblot (E), and qRT-PCR (F) analysis were performed using Jagged2 specific antibody and probe (**P < 0.01; Scale bars 100 μm). G Diagram of the mesothelium in 3D culture. Primary human omental fibroblasts are plated with human omental ECM and cultured for 6 hours. The fibroblasts are overlaid with human omental mesothelial cells and cultured for an additional 24 hours before OvCa cells are seeded. The purpose of the use of the 3D culture system was to investigate OvCa cells’ adhesion, invasion, and proliferation. H Ex-vivo human omentum adhesion, invasion, and proliferation assay with PHK25 labeled OvCa cells and the schematic of cell sorting by FACS. I Western blot analysis of Jagged2 protein in the surface cells of the human omentum in two culture conditions. Cells were cultured in human omentum without fluorescently labeled cells (unattached) or PHK25 labeled SKOV3-ip cells (attached) after FACS sorting. J qRT-PCR mRNA analysis of Jagged2 following the same methods described in H (**P < 0.01).