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Inhibition of EGFInduced ERK1 and ERK2 Enzyme Activation in NIH3T3 Cells by in situ...

Inhibition of EGFInduced ERK1 and ERK2 Enzyme Activation in NIH3T3 Cells by in situ Electroporation of a Nonpermeant Inhibitor

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Abstract

A nonpermeant compound, [(dimethylamino)methyl]acrylo-para- [(hydroxy-benzoylsulfonyl)-oxy]phenone, inferred by in vitro studies to specifically inhibit EGF receptor function, was introduced by in situ electroporation into mouse NIH3T3 fibroblasts growing on indium-tin oxide-coated glass. Cells were subsequently stimulated with growth factors and assessed for activation of a downstream target, the extracellular signal regulated kinase (ERK1/2), by probing with highly specific antibodies. The results showed that this compound can inhibit EGF- but not PDGF-mediated ERK1/2 activation in vivo, demonstrating the specificity of this compound and the utility of the in situ electroporation approach for the study of tyrosine kinase action in intact cells.

Promega Notes 67, 12.

Leda Raptis1, Heather L. Brownell1, Nicholas B. Lydon2 and Thomas M. Roberts3

1Queen's University, Ontario, Canada; 2Novartis, Switzerland; 3Dana-Farber Cancer Institute

Publication Date: 1998

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