Understanding the key features of the spontaneous formation of bona fide prions through a novel methodology that enables their swift and consistent generation

Table 1 Intracerebral inoculation of spontaneously generated PMSA products with distinct electrophoretic migration patterns in TgVole 1 × and TgVole 4 × animals

Strain	Passage	Substrate	TgVole (1 × ) mice
Strain	Passage	Substrate	Attack rate	Days post-inoculation (± SEM)	PrP^Sc classic pattern (WB)	Spongiosis	PrP^res (IHC)
Ust01^a	1st	rec-Vole	100%	156 ± 6	8/8	8/8	8/8
Ust02	1st	rec-Vole	100%	104 ± 3	8/8	8/8	8/8
Ust06	1st	rec-Vole	100%	201 ± 7	8/8	8/8	8/8
Ust09	1st	rec-Vole	100%	169 ± 5	7/7	7/7	7/7
TgVole-Ust01^b	2nd	Brain-TgVole	100%	94 ± 2	13/13	13/13	13/13
TgVole-Ust02	2nd	Brain-TgVole	100%	80 ± 3	6/6	6/6	6/6
TgVole-Ust06	2nd	Brain-TgVole	100%	100 ± 1	7/7	7/7	7/7
TgVole-Ust09	2nd	Brain-TgVole	100%	110 ± 2	5/5	5/5	5/5
TgVole (4×) mice
Ust02^a	1st	rec-Vole	100%	82 ± 1	9/9	9/9	9/9

^aRecombinant inoculum was prepared diluting the PMSA product 1:10 in PBS. 20 µl of the diluted inoculum were inoculated intracerebrally in each TgVole mouse
^bFor the second passage the brain homogenate of a diseased TgVole (selected for its incubation time close to the mean, and the intensity of its PrP^Sc signal in WB), was homogenized at 10% (w/V) in PBS (with protease inhibitor cocktail) and afterwards diluted 1:10 in PBS. 20 µl of the diluted inoculum were injected intracerebrally in each TgVole mouse

ISSN: 2051-5960