Fig 4. Latent HSV1 infection does not result in emergence of PrP misfolding in the E200K PrP organoids.

Organoids were infected with a 0.00001 MOI of McKrae HSV1 while treated with IFNα and brivudine from 24 hr before infection to 14 dpi. A. Example live organoid images of active infection at 3 dpi and latent infection at 23 dpi with or without reactivation by heat shock. Scale bar = 500 μm. B. Quantification of GFP signal following reactivation of HSV1 replication. Individual points show single organoids (n = 3) and horizontal lines show the average intensity. ***p<0.001, Student’s t-test. C. Protease resistant PrP detection in organoids at three months of latent infection compared with sCJD positive control brain homogenate. D. RT-QuIC seeding activity assays at 3 months post infection and E. at 6 months post-infection for the NKD donor. Traces shown are averages of 4 replicate reactions per ‘n’ individual organoids, with the ‘n’ indicated on each graph. None of the samples exceeded the 25% positive well threshold to be considered positive. F. Protease resistant PrP at 6 months of latent infection for the NKD donor compared with positive control sCJD homogenate (+). Numbers above lanes indicates individual organoids. Control organoids (age-matched, treated with compounds and ‘mock’ Vero cell lysates) can be found in S7 Fig.