Fig. 8

Effects of AhR/CYP1A1 activation and knockdown on β-Catenin levels in vitro and in vivo. a MCF-7 cells were treated with TCDD 10 nM and DMBA 5 μM for 72 h, thereafter the expression CYP1A1, β-Catenin, Cyclin D1 protein levels were determined by Western blot analysis using the enhanced chemiluminescence method and one of three representative experiments is shown. b MCF-7 cells treated with TCDD 10 nM were stained with primary antibodies against β-Catenin (green) followed by secondary antibodies and DAPI (red). Thereafter, β-Catenin localization and nuclear translocation was determined by immunofluorescence assay. Each sample was stained in triplicate for each antibody. c Twenty virgin female Balb/c mice were injected IP with either corn oil (vehicle) or single dose of DMBA 30 mg/kg. Two months later, mammary gland tissues were collected and stained with antibody against β-Catenin for IHC assay. d MCF-7 cells stably transfected with AhR shRNA (d) or CYP1A1 shRNA (e) were stained with primary antibodies against β-Catenin (green) and CYP1A1 (magenta) followed by secondary antibodies and DAPI (red). The β-Catenin and CYP1A1 localization and nuclear translocation were determined by immunofluorescence assay