Fig. 5.

Characterization of the AhR-KLF6 protein-protein interaction. Proteins generated as described in Fig. 4 were used in coimmunoprecipitation studies using lysates pretreated with vehicle (-) or 20 nM TCDD (+) for 20 minutes at 37°C. (A) Human KLF6 was immunoprecipitated and the lysates subjected to SDS-PAGE and Western blotting to detect coimmunoprecipitated hAhR. AhR and KLF6 protein were detected using the appropriate Cy3-labeled secondary antibodies (GE Healthcare), and images were captured using a Typhoon Trio Variable Mode Imager (GE Healthcare). (B) Reciprocal coimmunoprecipitation studies targeted the hAhR and assayed for coprecipitation of the hKLF6 proteins. (C) Murine KLF6 was used to coimmunoprecipitate the full-length mAhR as well as several deletion constructs. (D) Reciprocal coimmunoprecipitation used mAhR to pull down the mKLF6 deletion constructs.