Figure 2: Sensitive and specific prion seeding activity from organotypic slice cultures on RT-QuIC assay. | Scientific Reports

Figure 2: Sensitive and specific prion seeding activity from organotypic slice cultures on RT-QuIC assay.

From: Integrated Organotypic Slice Cultures and RT-QuIC (OSCAR) Assay: Implications for Translational Discovery in Protein Misfolding Diseases

Figure 2

(A) Slice cultures from WT mice (C57BL/6 background) efficiently seed the RT-QuIC assay. Representative RT-QuIC responses shown for the reactions seeded with 5 ng of WT homogenates from either NBH or RML scrapie-infected slice cultures (21 dpi) or brain homogenates from mock or terminally-infected scrapie mice. Uninfected samples did not show seeding activity and stayed at the baseline throughout the reaction. (B) RT-QuIC traces for a log dilution series from Tga20 slice cultures (21 DPI) showing clear concentration-dependent seeding activity. (Positive control = mock or clinical scrapie mice. Neg Con = mock/normal brain homogenate-infected samples). (C) Cultured slices from PrP−/− did not show seeding activity (both NBH and RML scrapie groups) at any seed concentration. Data from the traces shown are average of three replicates for each dilution. (D) Quantification of average ThT fluorescence values obtained over 60-hour period for each seed concentration tested from Tga20 slice cultures (21 dpi) shows a seed concentration-dependent increase. (E) Quantification of average amyloid formation rate (AFR) for each seed concentration tested from Tga20 slice cultures (21 dpi) showed a seed concentration-dependent increase in AFR.

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