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. 2019 Sep 11;9:13090. doi: 10.1038/s41598-019-49482-6

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© The Author(s) 2019

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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Downregulation of PrP^C upregulates α-SMA and fibronectin in the TM. (a) Probing of TM cell lysates treated with scrambled and PrP-siRNA for PrP^C shows the expected glycoforms in the scrambled control, and minimal reactivity for PrP^C in the experimental sample (lanes 1 & 2). (b) Probing of the same lysates for α-SMA and fibronectin shows upregulation in the absence of PrP^C relative to controls (lanes 1 & 2). (c) Quantification of protein expression by densitometry after normalization with β-actin shows 6.1-fold and 5.9-fold upregulation of α-SMA and fibronectin respectively. Values are mean ± SEM of the indicated n. *p < 0.05, **p < 0.01. Full-length blots (Supplementary Fig. S2). (d & e) Immunoreaction of fixed sections from the anterior segment of PrP^+/+ and PrP^−/− mice shows stronger reactivity for α-SMA and fibronectin in the TM of PrP^−/− relative to PrP^+/+ samples (panel 1 vs. 2). Scale bar: 25 µm. No reaction was detected in samples reacted with mouse IgG followed by Alexa 546-conjugated secondary antibody (panels 5 & 6).