Abstract

<p>. 10% weight/volume brain homogenates immunoblotted with rabbit polyclonal anti-human SOD1 (Abcam) and detected with IRDye800CW (green) conjugated goat anti-rabbit IgG (Li-Cor). Anti-β-actin mouse monoclonal antibody (Sigma) was also included as a loading control and was detected using IRDye680 (red) conjugated goat anti-mouse IgG (Li-Cor). Fluorescence was visualised using an Odyssey infra-red imager (Li-Cor). (A) Brains from <i>Sod1<sup>+/+</sup></i> wild type mice inoculated with PBS compared with end stage Sod1<sup>+/+</sup> wild type mice inoculated with RML, ME7 and MRC2 prion strains. No differences were seen between the groups. (B) Uninfected mice. (C) Total SOD enzymatic activity in 10% (w/v) <i>Sod1<sup>+/+</sup></i> (WT) brains. Brains from terminally sick mice infected with RML, ME7 and MRC2 were compared with uninfected mice. Samples were run in triplicate with n = 6 for each group. Data are shown normalised by total protein content (µg/ml) as determined by a Bradford protein assay (mean ± standard deviation). No significant difference was seen between the groups. (D) PrP<sup>c</sup> levels in <i>Sod1<sup>−/−</sup></i> and <i>Sod1<sup>+/+</sup></i> (WT) litter mate control mice by ELISA. PrP<sup>c</sup> levels (µg/ml) were determined in triplicate using 10% (weight/volume) brain homogenate for <i>Sod1<sup>−/−</sup></i> (n = 3) and <i>Sod1<sup>+/+</sup></i> (n = 3) in a PrP specific ELISA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054454#pone.0054454-Wadsworth2" target="_blank">[32]</a>. Data are shown normalised by total protein content (µg/ml and x 1000) as determined by a BCA assay (mean ± standard deviation). No significant difference was seen between the two groups.</p

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