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pp. High density reverse-phase protein lysate microarray Satoshi Nishizuka, Kazushige Ishida, Hironobu Noda, Masahiro Takahashi and Go Wakabayashi U.S. National Cancer Institute, Department of Surgery, Iwate Medical University School of Medicine, Uchimaru, Morioka, Iwate, Japan SDI Succinic dehydrogenase inhibition CCK- Cell Counting Kit ATP RPA Reversephase Protein Lysate Microarray RPA Paweletz RPA

Laemmli Pink SDS M urea SDS CHAPS BME mm DTT M TrisHCl glycerol bromphenol blue Pharmalyte ph - bromphenol blue SDS, sodium dodecyl sulfate; BME, beta mercaptoethanol; CHAPS, - -cholamidopropyl dimethylammonio - -propanesulfonate; DTT, dithiothreitol. RPA cm RPAHCT Pink buffer DTT SDS CHAPS V µl Pink buffer Aushon RPA pl µl

A semi-log scale T- Pink buffer T- HCT confluent µl RPA RPA

B RPA RPA single band at expected range no band relational database RPA RPA RPA TSA Tyramide Signal Amplification DAB -diaminobenzidine

C http: mttab.cancer.gov RPA perturbation RPA p -Mdm Gy γ p p Mdm p Mdm p Mdm

D RPA ASKMD A B C pd trastsumab RPA RPA RPA

p -Mdm A p Gy γ Mdm p Mdm B p Mdm p Mdm Brett Spurrier Sundhar Ramalingam Nishizuka, S., Ramalingam, S., Spurrier, B., Washburn, F.L., Krishna, R., Honkanen, P., Young, L., Shimura, S., Steeg, P.S., & Austin, J. J. Proteome Res.,,. Nishizuka, S. & Spurrier, B. Curr. Opin. Biotechnol.,,. Weinstein, J.N., Myers, T.G., O Connor, P.M., Friend, S.H., Fornace, A.J. Jr., Kohn, K.W., Fojo, T., Bates, S.E., Rubinstein, L.V., Anderson, N.L., Buolamwini, J.K., van Osdol, W. W., Monks, A.P., Scudiero, D.A., Sausville, E.A., Zaharevitz, D.W., Bunow, B., Viswanadhan, V.N., Johnson, G.S., Wittes, R.E., & Paull, K.D. Science,,. Yamori, T., Matsunaga, A., Sato, S., Yamazaki, K., Komi, A., Ishizu, K., Mita, I., Edatsugi, H., Matsuba, Y., Takezawa, K., Nakanishi, O., Kohno, H., Nakajima, Y., Komatsu, H., Andoh, T., & Tsuruo, T. Cancer Res.,,. Paweletz, C.P., Charboneau, L., Bichsel, V.E., Simone, N.L., Chen, T., Gillespie, J.W., Emmert-Buck, M.R., Roth, M.J., Petricoin III, E.F., & Liotta, L.A. Oncogene,,. Nishizuka, S., Charboneau, L., Young, L., Major, S., Reinhold, W.C., Waltham, M., Kouros-Mehr, H., Bussey, K.J., Lee, J.K., Espina, V., Munson, P.J., Petricoin, E. rd, Liotta, L.A., & Weinstein, J.N. Proc. Natl. Acad. Sci. U.S.A.,,. Nishizuka, S. Eur. J. Cancer,,. Ramalingam, S., Honkanen, P., Young, L., Shimura, T., Austin, J., Steeg, P.S., & Nishizuka, S. Cancer Res.,,. Spurrier, B., Honkanen, P., Holway, A., Kumamoto, K., Terashima, M., Takenoshita, S., Wakabayashi, G., Austin, J., & Nishizuka, S. Biotechnol. Adv.,,. Major, S.M., Nishizuka, S., Morita, D., Rowland, R., Sunshine, M., Shankavaram, U., Washburn, F., Asin, D., Kouros-Mehr, H., Kane, D., & Weinstein, J.N. BMC Bioinformatics,,. Spurrier, B., Washburn, F.L., Asin, S., Ramalingam, S., & Nishizuka, S. Proteomics,,. Engelman, J.A., Zejnullahu, K., Mitsudomi, T., Song. Y., Hy-

land, C., Park, J.O., Lindeman, N., Gale, C.M., Zhao, X., Christensen, J., Kosaka, T., Holmes, A.J., Rogers, A.M., Cappuzzo, F., Mok, T., Lee, C., Johnson, B.E., Cantley, L.C., & Jänne, P.A. Science,,. Kung, L.A. & Snyder, M. Nat. Rev. Mol. Cell. Biol.,,. Ma, H., Deacon, S., & Horiuchi, K. Expert Opin. Drug Discov.,,. Spurrier, B., Honkanen, P., Austin, J.,.