Figure 3: Temporal seeding activity from prion-infected organotypic slice cultures: RT-QuIC end-point dilution analysis in slice cultures from wild-type mice. | Scientific Reports

Figure 3: Temporal seeding activity from prion-infected organotypic slice cultures: RT-QuIC end-point dilution analysis in slice cultures from wild-type mice.

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

Figure 3

(A) Organotypic cerebellar slices from WT mice infected with scrapie show time-dependent seeding activity. After RT-QuIC reactions were seeded with 5 ng of homogenate, seeding activity occurred only in RML scrapie-infected slice cultures, not in NBH-exposed slice cultures. NBH-treated slices and RML scrapie-infected slices at 1 dpi did not show seeding activity. The earliest detection of seeding activity was observed at 7 dpi with longer lag phases. Seeding strength showed a clear time-dependent increase as judged by time to reach threshold. Seeding activity was also noted at 14 dpi, but the traces became stronger from slices at 21 dpi and continued to increase for later time points (28, 35 and 42 dpi). Data from each trace shown are the average ThT fluorescence of three replicate wells. (B–F) Seeded amplification of prions at log dilutions tested from cultured slices. Traces shown are average of 3 replicates from 42 dpi (B), 35 dpi (C), 21 dpi (D), 14 dpi (E) and 7 dpi (F).

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