Figure 6

Hotair epigenetically silences miR-568 in metastatic breast cancers. (A) qRT-PCR assays of Hotair expression in breast cancer cell lines. Relative levels compared with MCF-10A cells were plotted. (B) qRT-PCR (left) and transwell (right) assays of MDA-MB-231 cells following transfection with the indicated siRNAs. Relative levels compared with mock-transfected cells were plotted. (C) Clinical breast cancer samples were divided into Hotair-low and -high groups (left), and the probablities of survival were plotted based on a review of in-patient records and a follow-up study (right). (D) Pearson correlation analysis of the relative expression levels of Hotair (normalized to glyceraldehyde-3-phosphate dehydrogenase, GAPDH) and miR-568 (normalized to U6) determined using qRT-PCR in 29 human breast cancer tissue samples. (E) qRT-PCR assay of MDA-MB-231 cells transfected with siRNAs targeting the indicated genes. (F) ChIPs were performed using the indicated antibodies, followed by amplification of the 5′ regulatory region of miR-568. All data are representative photographs or represent the mean ± SD of three replicates. *P <0.05, **P <0.01 compared with all other groups.