Figure 7

Inhibition of Wnt/β-catenin signaling blocked the functional role of transducin (β)-like 1 X-linked receptor 1 (TBLR1). (A) Western blotting shows the protein levels of TBLR1, TCF4-dn, pRb and total Rb in breast cancer cell lines; β-tubulin was used as a loading control. (B) Indicated cells transfected with TOPflash or FOPflash and Renilla pRL-TK plasmids were subjected to dual-luciferase assays 48 hours after transfection. Reporter activity detected was normalized by Renilla luciferase activity. (C) PCR analysis shows the expression of cyclin D1, MYC and AXIN2 mRNA in breast cancer cell lines. Expression levels were normalized to glyceraldehyde-3-phosphate dehydrogenase. (D) Representative micrographs (left) and quantification (right) of crystal violet-stained cell colonies. (E) Anchorage-independent growth assays in indicated cells. (F) Flow cytometry analysis examined the portions of cell cycle in SKBR3 cells.