Journal: Molecular Neurodegeneration

Article Title: A Fluorescent Oligothiophene-Bis-Triazine ligand interacts with PrP fibrils and detects SDS-resistant oligomers in human prion diseases

doi: 10.1186/s13024-016-0074-7

Figure Lengend Snippet: RCA efficiently detects rSDS-PrP Sc oligomers in patients with sporadic Creutzfeldt-Jakob disease (sCJD). a Freshly homogenized human brain tissues from patients with sCJD (codon 129 Met/Met or Val/Val polymorphisms) were analyzed using RCA. The number on the figures corresponds to the identification numbers attributed by the tissue bank. The brain homogenate from a healthy control (NBH, the same as in Fig. ) was used as negative control. 50 μL of 10 % human brain homogenates in 100 μL PBS/2 % Sarkosyl were incubated with 2 mM MR100 at room temperature for 2 h and centrifuged at 11,000 g for 5 min. Supernatants (S) and pellets (P) were immunoblotted with the SAF mix. Molecular masses (20–75 kDa) are indicated on the left side of the panels. b The table below shows: ( i ) the duration (in months) of the symptomatic phase of the disease; ( ii ) the results of the densitometry analysis of oligomers after subtraction of the background to normalize each value (ImageJ software); and ( iii ) the results of the densitometry analysis of the monomers in the pellet (P) and in the supernatant (S), and the calculated ratio (P/S) obtained for each sample (Additional file : Table S1 for calculation of ratios). c The correlation between the oligomer amount and the duration of the symptomatic phase of the disease in the 10 sCJD samples was assessed with the Pearson’s test using GraphPad (San Diego, CA, USA)

Article Snippet: We used Pearson’s test with a probability of 0.05 (GraphPad software) and confirmed the presence of a significant correlation between duration of the symptomatic phase of the disease and intensity of the signal when all the ten sCJD samples were analyzed simultaneously (r 2 = 0.78, p-value = 0.0007) (Fig. ).

Techniques: Control, Negative Control, Incubation, Software