Fig. 2: Metformin treatment abolishes TCF4/β-catenin/CBP-p300 activation complex formation. | Cell Discovery

Fig. 2: Metformin treatment abolishes TCF4/β-catenin/CBP-p300 activation complex formation.

From: Cancer cell specific inhibition of Wnt/β-catenin signaling by forced intracellular acidification

Fig. 2

a Scheme of human SOX4 gene locus. Putative LEF/TCF binding sites marked with magenta boxes, and their positions in relation to the transcription start site (TSS, top arrow) are indicated. SOX4 gene consisting of a single exon is marked with a gray box; left and right thinner gray boxes mark positions of the 5’- and 3’-UTR regions, respectively. Small double arrows below the locus indicate positions of primer pairs used for ChIP analysis within the locus. bc ChIP assay done in H1299 cells treated with Wnt3a and Metformin, as indicated. After DNA recovery, binding efficiencies for TCF4 (b) or CBP and p300 (c) antibodies at 5 genomic locations spanning upstream, promoter (−800 and −400), exon and downstream regions of SOX4 gene, as well as a promoter of GAPDH and a known site for TCF4 binding at AXIN2 (b), - negative and positive TCF4 binding control genes, respectively, were measured by qPCR. Values were normalized to the total DNA input. Level of unspecific binding determined with normal IgG ChIP reactions are indicated with dashed black line. Error bars correspond to mean values ± SD. d Co-immunoprecipitation (Co-IP) assay done in H1299 treated as in (bc). After lysis, cells were subjected to immunoprecipitation with TCF4 antibody bound to agarose beads, and immunocomplexes were tested by WB with β-catenin antibody

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