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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: Cancer Res. 2015 Jan 29;75(7):1433–1444. doi: 10.1158/0008-5472.CAN-14-1026

Figure 5.

Figure 5

miR-135 and miR-203 directly target Runx2 in metastatic breast cancer cells. A, Luc-Runx2-3’UTR reporter functional assay. MiR-C, miR-135 and miR-203 were delivered in MDA-MB-231-luc cells and Firefly Luciferase activity was normalized to co-transfected Renilla luciferase and presented as fold change to miR-C. B, endogenous Runx2 protein was detected in miR-C, miR-135- and miR-203 transfected MDA-MB-231-luc cells by western blot analysis. C, Runx2 mRNA expression in primary tumors delivered miR-C, miR-135 and miR-203 was quantified by qRT-PCR. D, Runx2 protein in lung metastases after orthotopic (top) or intratibial tumor transplantation (lower) was analyzed by immunohistochemistry. E, expression of Runx2 target genes mediating osteolysis including IL-11, MMP-13 and PTHrP was analyzed in miR-C, miR-135 and miR-203 transfected cells (top) and in orthotopically transplanted mammary tumors (lower) by qRT-PCR. GAPDH was used as control. *significant difference (p < 0.05) between indicated miRNA and miR-C, Tukey’s post-hoc test. Scale bars 50 μm.