Observation of mechanisms for transcriptional regulation

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1.By the chemiluminescent or fluorescent immunohistochemistry, HSP90 localized with arylhydrocarbon receptor(AhR) was dissociated from AhR in the cytoplasm but not in the nuclei after the stimulation of 3 -methylcholanthrene in the H4IIE cell lines and hepatocytes in sections from rat livers. However, FLET analysis revealed that the both proteins associated with a loose connection or with interconnecting factor(s). We applied this technique to examine the intracellular localization of a novel protein that localized exclusively in the endocrine cells.2.Aclose relationship was found between the expression of CYP2B2 and area of endoplasmic reticulum(ER) in hepatocytes from normal and phenobarbital-treated rats as revealed by a combination of highly-sensitive and quantitative in situ hybridization of CYP2B2 mRNA with stereological analysis of ER.3.When a reporter gene was connected to 5'-region of CYP1A1 gene, we could detect the localization of the gene by the photon imaging. However, no significant difference was found in the DNA structure if 5 '-region of CYP1A1 gene and AhR-Arnt were reacted under the presence of transcriptional factors, Sp1 or CBP/p-300 by the photon detection and/or atomic-force microscopy.