Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

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

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Extended Data Fig. 2 — (a) Representative images of expression levels of Synapsin 1 (Syn1, upper) and Calbindin 1 (Calb1, lower) show predominant (Syn1) or almost exclusive (Calb1) expression in Purkinje cells (pc) in the cerebellar cortex. Image credit: Allen Institute. Scale bar = 100 µm. (b) Fluorescent micrographs of Prnp^ZH3/ZH3 COCS transduced with the AAV outlined in panel (A) show mNeonGreen expression predominantly in calbindin 1-expressing Purkinje cells. Scale bar = 50 µm. cgl = cerebellar internal granular layer, pc = Purkinje cell layer, ml = molecular layer. These findings were repeated in three independent experiments. (c) Left panel: Stably transfected CAD5-mPrP^C and CAD5-mPrP^R207A cells show similar PrP^C expression levels. epresentative PrP^C levels of one cell culture passage are shown. Right panel: POM19 immunoreactivity is divided by actin immunoreactivity, values are given as percentages of PrP^C. One datapoint corresponds to one passage of CAD5 cells. (d) Surface plasmon resonance (SPR) traces showing binding of POM1 to recombinant mPrP^R207A (rmPrP^R207A, k_a=3.8E⁺⁰⁵ 1/Ms, k_d=1.8E⁻⁰⁴ 1/s, K_D=4.7E⁻¹⁰ M; for comparison binding to recombinant wild-type murine PrP showed k_a=3.6E⁺⁰⁵ 1/Ms; k_d=9.1E⁻⁰⁵ 1/s; K_D=2.5E⁻¹⁰ M). (e) Immunohistochemistry of CAD5 Prnp^-/- cells stably transfected with pcDNA3.1 vector expressing wild-type murine PrP^C (mPrP^C), mPrP^R207A and mPrP^2cys. Monoclonal anti-PrP^C antibodies targeting distinct conformational epitopes on the globular domain of PrP^C were incubated to assess conformational changes in mPrP^R207A (POM1: α1-α3, POM5: β2-α2, POM8: α1-α2, POM19: β1-α3). Except for diminished staining of POM1 in mPrP^R207A, we observed robust detection of mPrP^R207A by POM5, POM8 and POM19 and mPrP^2cys by POM8 and POM19. Parts of this experiment, for example POM1 and POM19, were repeated twice. Scale bar = 20 µm.

Source data