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Furthermore, expression vectors for the fusion proteins were constructed and transfectedinto HeLa cells. Using the protein-overexpressed HeLa cell lysate, fluorescent andluminescent changes during incubation with Aβ were monitored. As the samples wereincubated for 50 hr, Y-Aβ-L luminescence intensity with Aβ1-42 was significantly moredecreased than that without Aβ1-42 (ca. 2 times), whereas there were little changes inY-Aβ-L fluorescence both with/without Aβ1-42. It was additionally found thatfluorescence intensities of thioflavin T with Aβ1-42 in cell lysate were increased dependingon the incubation time in the thioflavin T assay. These results indicated that the decrementof Y-Aβ-L luminescence denoted Aβ aggregation. Additionally, intra- or extracellularlocalization of aggregated Aβ was analyzed using the protein.Throughout these experiments, the detection system for Aβ localization andaggregation was achieved with luminescence changes providing Aβ conformationalinformation (Aβ aggregation information) and with fluorescence changes giving Aβlocalization information. With more improvements, this system would be one of thepowerful tools for the study of amyloidogenic protein behaviors and localization in internaland external cells.AcknowledgmentsThis study was in part supported by the Grants-in-Aid for Scientific Research, the "Core research"project (2009-2014) from the Ministry of Education, Culture, Sports, Science and Technology(MEXT). K.U. is grateful to Grant-in-Aid for Research Activity Start-up from MEXT. T.A. is gratefulfor Research Fellowships of the Japan Society for the Promotion of Science (JSPS) for YoungScientists.References1. Usui, K., Hulleman, J.D., Paulsson, J.F., Siegel, S.J., Powers, E.T., Kelly, J.W. Proc. Natl. Acad.Sci. U.S.A. 106, 18563-18568 (2009).2. Wurth, C., Guimard, N.K., Hecht, M.H. J. Mol. Biol. 319, 1279-1290 (2002).3. Kim, W., Hecht, M.H. Proc. Natl. Acad. Sci. U.S.A. 103, 15824-15829 (2006).489