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Inhibition
of EGF-Induced ERK1 and ERK2 Enzyme Activation in NIH3T3 Cells by in situ
Electroporation of a Nonpermeant Inhibitor
By Leda Raptis and Heather L. Brownell,
Queen's University, Kingston, Ontario, Canada, Nicholas B. Lydon, Novartis, Basel
Switzerland, and Thomas M. Roberts, Dana-Farber Cancer Institute, Boston, MA
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.
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