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. Author manuscript; available in PMC: 2013 Apr 20.
Published in final edited form as: Sci Transl Med. 2012 Aug 22;4(148):148ra117. doi: 10.1126/scitranslmed.3003808

Fig. 3.

Fig. 3

SAH-BCL9B inhibits Wnt transcriptional activity. (A) Colo320 cells were transfected with TOP-FLASH, incubated with vehicle or SAH-BCL9 peptides (10 μM), and assayed for luciferase activity, which was normalized to Renilla luciferase control. Error bars are mean +/− s.d. for assays performed in triplicate. *p < 0.01. (B) HCT116 cells that express low levels of BCL9 were transfected with TOP-FLASH and the indicated amounts of pcDNA-BCL9, treated with vehicle or SAH-BCL9 peptides (5 μM), and subjected to dual luciferase assay, performed at 24h. *p < 0.01. (C) Effect of SAH-BCL9B on NFκB transcriptional activity in HCT116 cells. (D) Effect of SAH-BCL9 peptides on dGFP expression in Colo320 cells lentivirally-transduced with a reporter containing TCF regulatory sequences (7xTdG). (E) ChIP of Colo320 cells using anti-TCF-4, β-catenin, and BCL9 antibodies. Negative controls included ChIP with IgG and the use of primers to a non-specific, upstream region of the VEGF promoter. (F) Colo320 cells lentivirally-transduced with control or BCL9 shRNA vectors were transfected with VEGF promoter-luciferase reporter plasmids. Reporter activity was assayed using the dual luciferase assay system and results normalized to Renilla values for each sample. *, p< 0.001. (G) qRT-PCR analysis of Wnt target gene expression in response to SAH-BCL9B treatment of Colo320 cells. Error bars are mean +/− s.d. for assays performed in quadruplicate. * p < 0.01.