PK-resistant PrP in light fractions of TgMHu2ME199K brain gradients.

<p>Sarkosyl extracted brain homogenates of sick TgMHu2ME199K/wt mice (in the figure: 9 month old score = 4) were subjected to ultracentrifugation in 10–60% sucrose gradients. Individual fractions were digested in the presence or absence of PK and immunoblotted with: (<b>a</b>) α-PrP mAb 6H4 and (<b>b</b>) α-PrP pAb RTC directed against the C-terminal end of PrP.</p

PrP forms in soluble and membranal brain fractions.

<p>(<b>a</b>) Fractionation protocol of brain homogenates. (<b>b</b>) Soluble and membranal brain fractions were digested in the presence and absence of PK and immunoblotted with two α-PrP c-terminal Ab; pAb RTC and mAb EP1802Y (EP) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0069583#pone-0069583-g002" target="_blank">figure 2a</a> for epitope mapping).</p

Similar disease kinetics and PrP accumulation in heterozygous and homozygous TgMH2ME199K mice.

<p>(<b>a</b>) The percentage of mice presenting a score <2 in both E199K/ko and E199K/wt lines as related to age. Median was 5.2±0.8 for TgMHu2ME199K/ko mice and 6.0±1.2 for TgMHu2ME199K/wt mice. (<b>b</b>) PK-resistant PrP levels are similar in both lines as related to age. Brain homogenates from both TgMHu2ME199K/KO and TgMHu2ME199K/wt lines in different ages and clinical stages (1 month, 3 months and 7 months, score = 0, score = 0 score = 3 respectively) as well as wt and scrapie RML controls were digested with PK and immunoblotted with α-PrP pAb RTC.</p

Correction: PrP^ST, a Soluble, Protease Resistant and Truncated PrP Form Features in the Pathogenesis of a Genetic Prion Disease

<p>Correction: PrP^ST, a Soluble, Protease Resistant and Truncated PrP Form Features in the Pathogenesis of a Genetic Prion Disease</p

Aggregation of wt and mutant PrP in TgMHu2ME199K/wt mice.

<p>(<b>a</b>) <b>Epitope mapping of α-PrP antibodies:</b> epitopes of antibodies used in this manuscript are depicted on a schematic representation of the chimeric mouse-human E199K PrP. In the next panels (b &c) αPrP mAbs IPC1 and 3F4 were used to differentiate between wt PrP and chimeric-mutant PrP respectively. (<b>b</b>) <b>Oligomeric E199K PrP in asymptomatic TgMHu2ME199K/ko mice:</b> Sarkosyl extracted brain homogenates of wt and 3 months old (score = 0) TgMHu2ME199K/ko mouse were subjected to ultracentrifugation in 10–60% sucrose gradients <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0069583#pone.0069583-Tzaban1" target="_blank">[34]</a>. Individual fractions were immunoblotted with αPrP mAb IPC1 to detect wt PrP and αPrP mAb 3F4 to detect chimeric-mutant PrP. (<b>c</b>) <b>Oligomeric wt PrP in sick TgMHu2ME199K/wt mice.</b> Sarkosyl extracted brain homogenates from TgMHu2ME199K/wt mice at different ages (1 month, 3 months and 7 months, score = 0, score = 0 score = 3 respectively) were subjected to ultracentrifugation in 10–60% sucrose gradients. Individual fractions of each gradient were immunoblotted with mAb IPC1 to detect wt PrP and mAb 3F4 to detect chimeric-mutant PrP.</p

Correction: Fatal Prion Disease in a Mouse Model of Genetic E200K Creutzfeldt-Jakob Disease

<p>Correction: Fatal Prion Disease in a Mouse Model of Genetic E200K Creutzfeldt-Jakob Disease</p