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Cover each source plate containing small molecule library with an aluminum sealing tape. Ensure that the tape forms a tight seal using a rolling sealer. Put the source plates back in a –80°C freezer for long-term storage. 4. Screening for Effects of Small Molecules by Visual Inspection of Phenotypes 1. ” 2. At desired times in development, remove the 96-well plates containing compound-treated embryos from the incubator and examine each well under a stereomicroscope. For better visualization of subtle changes, such as changes in circulatory pattern, a phase-contrast inverted microscope can be used.

High-throughput cell-based assays have proven to be powerful tools to deconvolute complex signaling pathways, including the MAP kinase pathways. For many decades, in vitro culture of mammalian cells has been harnessed to study basic biological questions. Diverse transformed and primary mammalian cells have been exploited to study diseases, including diabetes, cancer, and neurodegenerative disease (7, 8). However, many breakthroughs have depended on the serendipitous discovery of a small-molecule inhibitor with specificity for a particular biological process.

And Pour, P. M. (2002) Pancreatic cell lines: A review. Pancreas 24, 111–120. 9. , Yamamoto, T. and Nishida, E. (2004) Regulatory mechanisms and function of ERK MAP kinases. J. Biochem. (Tokyo) 136, 557–561. 10. Yoon, S. and Seger, R. (2006) The extracellular signal-regulated kinase: Multiple substrates regulate diverse cellular functions. Growth Factors 24, 21–44. 41 11. Gimbrone Jr, M. , Cotran, R. S. and Folkman, J. (1974) Human vascular endothelial cells in culture: Growth and DNA synthesis.

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