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Additional Supporting Information may be found in the online version of this article.

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sc-12-0190_sm_SupplFigure1.tif249KSupplemental Figure 1. Characterization of human ovarian cancer stem-like cells derived from an ovarian cancer cell line. A. Representative phase-contrast photomicrographs of A2780 cells and spheres cultured under stem cell-conditions; from left to right: A2780 cells, passage 1, passage 2, passage 3, and passage 4 cells, respectively. B. Flow cytometric analysis of CD133 expression in A2780 cells, passage 1, passage 2, passage 3, and passage 4 cells, respectively (from left to right). C. RT-PCR analysis of Oct-4, Nanog, and CD133 mRNA expression in A2780 cells, CSLCs, and CSLCs cultured under differentiating conditions for 11 days; GAPDH mRNA expression is used as a control. D. Representative staining of spheroids for CD133 expression by immunofluorescence; nuclei were counterstained with DAPI. E. Adherence of dissociated sphere-forming cells to plates after culture under differentiating conditions for 11 days; magnification: 100×(right) and 200×(medium), and immunohistochemical staining of CA125 in differentiated cells (left). F. Tumorigenic potential of 1×104 CSLCs (CD133-positive) compared with 1×105 CD133-negative cells (A2780). Cells were simultaneously injected into the right and left flank of the same mouse and tumor formation assessed after 5 weeks.
sc-12-0190_sm_SupplFigure2.tif212KSupplemental Figure 2. Characterization of human ovarian cancer stem-like cells isolated from primary ovarian cancer tissues. A. Representative staining of spheroids for CD133 expression by immunofluorescence; nuclei were counterstained with DAPI. B. Representative phase-contrast photomicrographs of spheres, isolated from primary ovarian cancer tissues, cultured under stem cell-conditions (left). After culture under differentiating conditions for 11 days, dissociated sphere-forming cells adhere to plates (right). Magnification 200×. and immunohistochemical staining of CA125 in differentiated cells C. Flow cytometric analysis of CD133 expression in primary cancer cells (left), and in CSLCs isolated by magnetic activated cell sorting (right). D. Tumorigenic potential of 1×104 CSLCs (CD133-positive) compared with 1×105 primary cancer cells (CD133-negative). Cells were simultaneously injected into the right and left flank of the same mouse and tumor formation assessed after 5 weeks.
sc-12-0190_sm_SupplFigure3.tif1406KSupplemental Figure 3. Chemokine and chemokine receptor gene expression levels in ovarian CSLCs. A. Relative expression levels (fold-change) of chemokines and chemokine receptors down-regulated (-ve numbers) or up-regulated (+ve numbers) in CD133-positive CSLCs vs CD133-negative NCSLCs, derived from the A2780 cell line, as measured by PCR array. B. Gene expression levels of CXCL16 and its receptor CXCR6 as determined by qRT-PCR. C. Protein expression levels of CXCL16 protein as measured by ELISA, was performed in CD133-positive and negative cells, isolated from three ovarian cancer patient and A2780 cells. D, CSLCs derived from A2780 cells, were plated on the upper cell culture inserts, with various concentration of anti-CXCL16 antibody placed in the lower chambers. In vitro migration were measured by transwell migration assays after 24h. Bars correspond to mean ±SD. *P<0.05, **P<0.01.
sc-12-0190_sm_SupplFigure4.tif1731KSupplemental Figure 4. CD133-positive CSLCs transfected with control shRNA or CCL5-shRNA. A. Representative phase-contrast photomicrographs (left panels) and fluorescence photomicrographs (right panels) of CD133-positive cells transduced with vector control shRNA or CCL5-shRNA. B. Flow cytometric analysis of GFP expression in CD133-positive cells transduced with vector control shRNA or CCL5-shRNA. C CCL5 gene expression levels in CD133-positive cells transduced with vector control shRNA or CCL5-shRNA as determined by RT-PCR.. D. CCL5 protein expression levels, as measured by ELISA, in CD133-positive cells transduced with vector control shRNA or CCL5-shRNA. E. Cell proliferation assay was performed for CD133+ CSLCs transduced with vector control shRNA or CCL5-shRNA. Bars correspond to mean ±SD. *P<0.05, NS means no significant difference.
sc-12-0190_sm_SupplTable1.pdf8KSupplemental Table 1.
sc-12-0190_sm_SupplTable2.pdf75KSupplemental Table 2.
sc-12-0190_sm_SupplTable3.pdf78KSupplemental Table 3.

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