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Two types of prion-infected mouse neuroblastoma (N2a) cell lines were used in this study: N2a cells infected with the RML strain (ScN2a) [16] and N2a#58 cells infected with the Fukuoka-1 strain (F3). N2a#58 cells are known to express five times more normal PrP than N2a cells. Both ScN2a cells and F3 cells were grown in six-well culture plates in Opti-MEM (Invitrogen) supplemented with 10% fetal bovine serum. The cells were allowed to reach confluence, and chemicals at various concentrations were added to the medium when 5% of the confluent cells were passaged.The final concentration of either DMSO or ethanol in the medium was less than 0.2%. The cultures were allowed to grow to confluence (3 or 4 days).
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18,19 Solutions of 10 mM Cu(II) and each compound at a compound:Cu (II) ratio of 1:0 to 0:1 were prepared in ultrapure water (MilliQ; Millipore Co., Japan) or 95% ethanol, and λmax of the copper complex was measured.
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