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Figure 1 | Molecular Cancer

Figure 1

From: Rac1 GTPase and the Rac1 exchange factor Tiam1 associate with Wnt-responsive promoters to enhance beta-catenin/TCF-dependent transcription in colorectal cancer cells

Figure 1

Active Rac1 associates with Wnt-responsive promoters in vivo . (A) HCT116 cells with stable inducible expression of myc-tagged V12Rac1 were treated with (+) or without (-) 0.1 μg/ml doxycyline (dox) for 24 hours. Nuclear extracts were immunoprecipitated (IP) with myc-tag or β-catenin antibodies and immunocomplexes were analyzed by immunoblotting (IB) with β-catenin and myc-tag antibodies. Nuclear protein (20 μg) was loaded to confirm exogenous expression of myc-tagged V12Rac1 and endogenous β-catenin expression. Topoisomerase II and Paxillin were used as markers for nuclear and cytoplasmic fractions, respectively. (B) HCT116 cells with stable inducible expression of myc-tagged V12Rac1 were treated with (+) or without (-) 0.1 μg/ml doxycycline (dox) for 24 hours. Chromatin was isolated from formaldehyde-fixed cells, and subjected to immunoprecipitation with TCF-4, β-catenin, Rac1, or myc-tag antibodies. IgG or no antibody were used as negative controls. Enriched chromatin was PCR amplified using primers that flank TBEs in c-Myc or Cyclin D1 promoters. As a negative control, primers specific to the GAPDH promoter, which is devoid of TBEs, were used. Input represents 10% of chromatin used for immunoprecipitation.

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Molecular Cancer

ISSN: 1476-4598