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. 2022 Aug 10;29(8):831–840. doi: 10.1038/s41594-022-00814-7

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a, Representative magnetic resonance diffusion-weighted images (DWI) 24 hours after stereotactic injection of holo-^hcY104A (left). Contralateral injections of holo-^hcY104A + rmPrP₂₃₂₃₁ (right). A small area of hyperintensity was found in one mouse after injection of 12 µg holo-^hcY104A (white arrowhead). White asterisks: needle tract. b, Hematoxylin and eosin (HE)-stained sections from mice shown in a. Asterisks: needle tract. Rectangles denote regions magnified in c. c, HE sections (CA4). Left, holo-^hcY104A injections (6, 9 and 12 µg). Right, holo-^hcY104A + rmPrP_23–230. Asterisk (9 µg): neurons with hypereosinophilic cytoplasm and nuclear condensation in the vicinity of the needle tract. Asterisk (12 µg): These neurons were diffusely distributed among numerous healthy neurons. White arrowhead: vacuoles indicative of edema along the needle tract. d, DWI images of 6 µg holo-POM1 ± rmPrP_23–231, revealing a hyperintense signal at 24 hours. e, HE-stained section from a mouse shown in d. Asterisks: needle tract. Rectangles: areas in f. f, HE sections (CA4). Holo-POM1 injections revealed damaged neurons with condensed chromatin and hypereosinophilc cytoplasm. g, Volumetric quantification of lesions on DWI imaging 24 hours after injection revealed no significant lesion induction by holo-^hcY104A. One datapoint corresponds to an animal. P values are adjusted for multiple comparisons. n.s.: not significant, P > 0.05, ordinary one-way ANOVA with Šídák’s multiple comparisons test. h, Antibody expression levels, as determined by Myc-Tag western blot, showed a positive correlation with survival. One datapoint corresponds to one animal. Pearson correlation coefficient r = 0.72, 95% confidence interval 0.099–0.94, P = 0.03. a.u., arbitrary units. i, Significant correlation of PrP^Sc and antibody expression levels (representative images depicted in j, aggregated correlation across all brain regions). Different colors represent 3 brain regions from 9 independent animals. Pearson correlation coefficient r = 0.53, 95% confidence interval 0.18–0.76, P = 0.0048. j, Representative images from quantification of l. l, Sagittal brain sections stained with SAF84, highlighting PrP^Sc, and basal ganglia immunofluorescent micrographs marking ^hcY104A-Myc-tag. Scale bar SAF84: 1 mm. Scale bar ^hcY104A-Myc-tag: 500 µm.

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