Fig. 8: IL-6 secreted from omental mesothelial cells induce stemness in a Notch-Hes1 dependent fashion and stimulates the growth of Jagged2 expressing cells. | Cell Death & Disease

Fig. 8: IL-6 secreted from omental mesothelial cells induce stemness in a Notch-Hes1 dependent fashion and stimulates the growth of Jagged2 expressing cells.

From: Ovarian tumor cell-derived JAGGED2 promotes omental metastasis through stimulating the Notch signaling pathway in the mesothelial cells

Fig. 8

A A List of genes with an expression fold change between pleural malignant and peritoneal malignant mesothelial cells of more than 2-fold from the public dataset microarray GSE63966. B qRT-PCR mRNA analysis of the Notch target genes, and CTGF and FN1 from mesothelial cells, omental metastatic tumor cells, and primary ovary tumor cells resected from the patient. C (Left) Schematic model of the co-culture system of mesothelial and Jag2OE cells. (Right) qRT-PCR mRNA expression of the indicated genes in control and Jag2OE tumor cells cocultured with mesothelial cells. *P < 0.05, **P < 0.01, ***P < 0.001. D Heat map showing qRT-PCR mRNA expression levels of the indicated genes from mesothelial cells that were FACS separated from cocultures of each experimental/treatment group. E qRT-PCR analysis of Hes1 expression in mesothelial cells treated with scrambled or Hes1 siRNA and cultured in 24 well plates coated with either Fc or control or recombinant Jagged2 protein (n = 3). Data represent average +/- SD. **P < 0.01, ***P < 0.001. F Quantification of cell proliferation in mesothelial cells treated with control or Hes1 siRNA and cultured in 24 well plates coated with either Fc or control or recombinant Jagged2 protein by luciferase assay (n = 3). **P < 0.01, ***P < 0.001. G Quantification of IL-6 levels in the conditioned media of control or Jag2OE tumor cells cultured alone or cocultured with mesothelial cells in the presence of DMSO, MRK-003 (1μM) by ELISA (n = 3). **P < 0.01, ****P < 0.001. H Quantification of IL-6 levels in the conditioned media of the indicated tumor cells co-cultured with mesothelial cells after treatment with Hes1 siRNA by ELISA. ***P < 0.001. I Cell proliferation of the indicated tumor cells co-cultured with mesothelial cells from each experimental group by luciferase assay. Data represent average +/-SD, **P < 0.01, ***P < 0.001. J Cell proliferation of the indicated tumor cells cocultured with mesothelial cells after treatment with Hes1 siRNA by luciferase assay. **P < 0.01, ***P < 0.001. K Quantification of sphere formation capacity of the indicated tumor cells cocultured with mesothelial cells after treatment with Hes1 siRNA. **P < 0.01, ***P < 0.001. L Quantification of sphere formation capacity of indicated tumor cells cocultured with mesothelial cells after treatment with 1 μM MRK-003. **P < 0.01, ***P < 0.001. M Cell proliferation of indicated tumor cells cocultured with mesothelial cells and treatment with either IgG or 5.0 μg/mL of the anti-IL-6 antibody by luciferase assay. **P < 0.01, ***P < 0.001. N Cell proliferation of indicated tumor cells cocultured with mesothelial cells and treatment with either PBS, 5.0 μg/mL, or 10.0 μg/mL of anti-IL6 by luciferase assay. *P < 0.05, ***P < 0.001. Data in the figure represent average and +/-SEM; P-values were determined using the Student’s t-test unless otherwise indicated. All experiments were run in triplicates.

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