Cellular localization of induced human interferon-? mRNA by non-radioactive in situ hybridization

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Abstract
Induced interferon-β (IFN-β) mRNA was localized in human FS-4 fibroblasts by in situ hybridization using biotinylated probes. The hybridization sites were detected by incubation with a nick-translated genomic DNA probe (1.8 kb) via streptavidin-colloidal gold followed by silver contrast enhancement. The positive signals were observed by reflection-contrast light microscopy. IFN-β mRNA was transiently induced by poly r(I):r(C) in fibroblasts 2–4 h after induction. Induction in the presence of cycloheximide and actinomycin D (superinduction conditions) exhibited an enhanced level of IFN-β mRNA with a maximum at 4–8 h. The kinetics of the IFN-β mRNA expression in the cytoplasm as revealed by in situ hybridization proved to be compatible with the results of Northern biotting experiments of total cellular RNA.

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