Induction of human interferon-gamma mRNA analyzed by cytoplasmic dot hybridization: effect of Con A, TPA, and A23187.
The cytoplasmic dot hybridization method was improved by eliminating non-specific false signals. The non-specific binding to a probe disappeared almost completely when the hybridized nitrocellulose sheet was treated with pronase. Using this improved method we demonstrated that reagents such as 12-O-tetradecanoylphorbol-13-acetate (TPA), concanavalin A (Con A) and a calcium ionophore (A23187) could induce the expression of IFN-gamma gene of a human T-lymphoblastoid cell line, TCL-Fuj 2M, where TPA functioned synergistically with Con A and A23187. However, when the cells were cultured with the ionophore for more than one day the amount of IFN-gamma mRNA was markedly reduced to a low level.
D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D003208:Concanavalin A / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D012333:RNA, Messenger; D003208:Concanavalin A; D000001:Calcimycin; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate