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. Author manuscript; available in PMC: 2008 Mar 7.
Published in final edited form as: Toxicology. 2006 Dec 22;231(2-3):224–233. doi: 10.1016/j.tox.2006.12.019

Figure 6. EMSA of Caco-2 nuclear extract and super shift assay of hCAR and hRXRα with the IR2-containing oligonucleotide (31 bp).

Figure 6

Figure 6A, gel shift assay of Caco-2 nuclear extract with the IR2-containing oligonucleotide. The labeled IR2 wild type oligonucleotide (lane 1 – 4) or labeled mutated IR2 oligonucleotide (lane 5) was incubated with Caco-2 nuclear extract (lane 2–5, lane 1 for control without nuclear extract) for 20 minutes at room temperature to form possible DNA protein complexes. 125-fold excess of wild type cold competitors (lane 3) and mutated cold competitors (lane 4) was added to test the specificity of the IR2. Figure 6B, super shift assay of hCAR and hRXR. Antibody specific to hCAR (lane 6) and hRXRα (lane 7) was added to the nuclear extract and incubated for 20 minutes at room temperature, then followed with the shift reaction.