Co-localization of the nApoECF antibody with caspase-cleaved tau.

<p>(<b>A–C</b>): Representative immunofluorescence double-labeling in Case #4 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080180#pone-0080180-t001" target="_blank">Table 1</a>) within the CA1 region of the hippocampus in Pick's disease utilizing the nApoECF antibody (green, Panel A), caspase-cleaved tau (red, Panel B) and with the overlap image shown in Panel C. All scale bars represent 10 µm. (<b>D</b>): Quantification of Pick bodies double-labeled by TauC3 (caspase-cleaved tau) and nApoECF in area CA1 of the hippocampus. Data show the number of Pick bodies labeled with nApoECF alone (blue bar), Tauc3 alone (green bar) or those Pick bodies that were labeled with both antibodies (red bar). Pick bodies were identified in a 20× field within area CA1 by immunofluorescence overlap microscopy (n = 3 fields for 4 different Pick cases) ±S.D., *p<0.05. Data indicated that roughly 75% of all labeled Pick bodies within area CA1 were co-localized with both antibodies. (<b>E–G</b>): Double-labeled confocal immunofluorescence images with the nApoECF antibody (green, E), PHF-1 (red, F), and the two images overlapped (G). (<b>H</b>): High magnification confocal overlapped image representing nApoECF (green channel, left), TauC3 (red channel, middle), and overlapped image (yellow/orange, right). Scale bars in Panel H represent 5 µm.</p

Case Demographics.

<p>PMI, postmortem interval in hours; NPD, neuropathological diagnosis.</p

Labeling of nApoECF antibody within the hippocampus co-localizes with PHF-1 and is regionally defined.

<p>(<b>A–C</b>): Representative immunofluorescence double-labeling in Case #3 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080180#pone-0080180-t001" target="_blank">Table 1</a>) utilizing the nApoECF antibody (green, Panel A) and PHF-1 (red, Panel B) revealed strong co-localization of the two antibodies within Pick bodies of area CA1 (Panel C). (<b>D–F</b>): In contrast to area CA1, strong immunofluorescence was detected using PHF-1 (red, Panel E), but only weak labeling was observed within granule cells of the dentate gyrus following application of the nApoECF antibody (green, Panel D). Panel F shows the overlap image with yellow indicating Pick bodies double-labeled with both antibodies. (<b>G</b>): Quantification of Pick bodies double-labeled by PHF-1 and nApoECF. Data show the number of Pick bodies labeled with nApoECF alone (blue bar), PHF-1 alone (green bar) or those Pick bodies that were labeled with both antibodies (red bar) in area CA1. Pick bodies were identified in a 20× field within area CA1 by immunofluorescence overlap microscopy (n = 3 fields for 4 different Pick cases) ±S.D., *p<0.05. Data indicated that roughly 86% of all identified Pick bodies within area CA1 were labeled with both antibodies. (<b>H–J</b>): Double-labeled confocal immunofluorescence images with the nApoECF antibody (green, H), PHF-1 (red, I), and the two images overlapped (J). All scale bars represent 10 µm, except for confocal images in Panel H–J, which represent 50 µm. (<b>K</b>): High magnification confocal overlapped image representing nApoECF (green channel, left), PHF-1 (red channel, middle), and overlapped image (yellow/orange, right); notice filamentous nature of staining within the two Pick bodies. Scale bars for Panel K represent 5 µm.</p