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                <term xml:lang="en">image recognition</term>
                <term xml:lang="en">automation</term>
                <term xml:lang="en">high-content screening</term>
                <term xml:lang="en">second messenger-activated kinases</term>
                <term xml:lang="en">signal transduction</term>
                <term xml:lang="en">nucleocytoplasmic transport</term>
                <term xml:lang="en">NF-κB</term>
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              <p>Transcription factors of the nuclear factor kappa B family are the paradigm for signaling dependent nuclear translocation and are ideally suited to analysis through image-based chemical genetic screening. The authors describe combining high-content image analysis with a compound screen to identify compounds affecting either nuclear import or export. Validation in silico and in vitro determined an EC(50) for the nuclear export blocker leptomycin B of 2.4 ng/mL (4.4 nM). The method demonstrated high selectivity (Z' &gt;0.95), speed, and robustness in a screen of a compound collection. It identified the IkappaB protein kinase inhibitor BAY 11 7082 as an import inhibitor, the p38 mitogen-activated protein (MAP) kinase inhibitor PD98509 as an import enhancer, and phorbol ester as an export inhibitor. The results establish a robust method for identifying compounds regulating nucleocytoplasmic import or export and also implicate MAP kinases in nuclear import of nuclear factor kappa B.</p>
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