Figure 5

Effects of TCDD on the transcription rate of vitellogenin, estrogen receptor α and cytochrome P4501A mRNAs. Nuclear run-off experiments using primary cultured hepatocytes treated with E2 (10 nM) or E2 (10 nM) plus TCDD (10 nM) for 8 hours. The cells were harvested and the activated nuclei were prepared. The [α-³²P]UTP incorporated total RNA was prepared by in vitro transcription assay as described in the methods. The radioactive labelled RNA samples were hybridized to the specific cDNAs for vitellogenin (VTG), estrogen receptor alpha (ERα) and cytochrome P4501A (CYP1A) cross-linked to the nylon membranes. Non-specific hybridization is indicated by hybridization of the labelled RNA with cloning plasmid pGEM-3Zf. The results shown are from a representative experiment repeated three times.