Abstract
The electrophoretic mobility shift assays (EMSA) with the use of the synthetic HIV-1 NF-kappa B motif as a probe, showed that LPS-treatment of J774 cells (a mouse macrophage cell line) leads to the activation of the fast-moving (denoted as B1) and the slow-moving NF-kappa B (denoted as B2). The binding of both B1 and B2 to the NF-kappa B probe was inhibited specifically by either unlabelled NF-kappa B, or competitor probes, but not by unrelated probes. LPS-induced activation of NF-kappa B was inhibited by a protein kinase A (PKA) inhibitor (H-89), but not by a protein kinase C (PKC) inhibitor (H-7). PMA itself failed to activate NF-kappa B and the depletion of PKC did not prevent LPS-induced activation of NF-kappa B. The pre-treatment of J774 cells with dibutyric cAMP, forskolin, prostaglandin E2 or cholera toxin resulted in NF-kappa B activation. Thus, these data suggested a probable involvement of PKA in LPS-induced NF-kappa B activation in macrophages.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
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Amino Acid Sequence
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Animals
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Base Sequence
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Bucladesine / pharmacology
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Cholera Toxin / pharmacology
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Colforsin / pharmacology
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Consensus Sequence
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Cyclic AMP / physiology*
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Dinoprostone / pharmacology
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Female
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Gene Expression Regulation, Neoplastic / drug effects
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Isoquinolines / pharmacology
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Lipopolysaccharides / antagonists & inhibitors
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Lipopolysaccharides / pharmacology*
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Macrophage Activation / drug effects*
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Macrophages / drug effects*
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Macrophages / enzymology
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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NF-kappa B / metabolism*
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Neoplasm Proteins / metabolism
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Phosphorylation
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Piperazines / pharmacology
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Protein Kinases / physiology*
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Protein Processing, Post-Translational
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Proto-Oncogene Proteins / metabolism
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Signal Transduction* / drug effects
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Sulfonamides*
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Tetradecanoylphorbol Acetate / pharmacology
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Transcription Factor RelB
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Transcription Factors*
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Transcription, Genetic* / drug effects
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Tumor Cells, Cultured
Substances
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Isoquinolines
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Lipopolysaccharides
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NF-kappa B
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Neoplasm Proteins
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Piperazines
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Proto-Oncogene Proteins
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Relb protein, mouse
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Sulfonamides
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Transcription Factors
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Transcription Factor RelB
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Colforsin
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Bucladesine
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1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
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Cholera Toxin
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Cyclic AMP
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Protein Kinases
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Dinoprostone
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N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
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Tetradecanoylphorbol Acetate