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. 2011 Apr 29;6(4):e19338. doi: 10.1371/journal.pone.0019338

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Masliah et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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To examine the effects of immunization on α-syn accumulation, immunohistochemical analysis using antibodies against FL-α-syn (layers 5–6) and CC-α-syn was conducted. Panels illustrate laser scanning confocal images of the temporal cortex and hippocampus (CA3) immunolabeled with antibodies against FL and CC α-syn immunoreactivity. (A, C) Temporal cortex of IgG1-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against FL-α-syn, respectively. (B, D) Temporal cortex of 9E4-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against FL-α-syn, respectively. (E, G) Hippocampus of IgG1-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against FL-α-syn, respectively. (F, H) Hippocampus of 9E4-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against FL-α-syn, respectively. (I) Image analysis of the numbers of neocortical α-syn immunoreactivity neurons with the FL α-syn antibody. (J) Analysis of the levels of α-syn immunoreactivity in the neuropil in the neocortex in sections labeled with the FL α-syn antibody. (K) Image analysis of the numbers of hippocampal α-syn immunoreactivity neurons with the FL α-syn antibody. (L) Analysis of the levels of α-syn immunoreactivity in the neuropil in the hippocampus in sections labeled with the FL α-syn antibody. (M, O) Temporal cortex of IgG1-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against CC α-syn, respectively. (N, P) Temporal cortex of 9E4-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against CC α-syn, respectively. (Q, S) Hippocampus of IgG1-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against CC α-syn, respectively. (R, T) Hippocampus of 9E4-immunized non-tg and α-syn tg mouse immunolabeled with an antibody against CC α-syn, respectively. (U) Image analysis of the numbers of neocortical α-syn immunoreactivity neurons with the CC α-syn antibody. (V) Analysis of the levels of α-syn immunoreactivity in the neuropil in the neocortex in sections labeled with the CC α-syn antibody. (W) Image analysis of the numbers of hippocampal α-syn immunoreactivity neurons with the CC α-syn antibody. (X) Analysis of the levels of α-syn immunoreactivity in the neuropil in the hippocampus in sections labeled with the CC α-syn antibody. Scale bar = 30 µM. N = 20 mice per group; 12 month old. Error bars represent mean ± SEM. (*) indicates p<0.05, when comparing IgG1-immunized α-syn tg mice to IgG1-imunized non-tg mice by one-way ANOVA with post hoc Dunnett's. (#) Indicates p<0.05, when comparing α-syn tg mice immunized the 9E4 α-syn antibody to IgG1-treated α-syn tg mice by one-way ANOVA with post hoc Dunnett's.