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. 2021 Sep 2;11:17557. doi: 10.1038/s41598-021-96181-2

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Over-expression of PDI is protective against inhibition of nuclear import induced by mutant FUS in a neuronal cell line. (A) Fluorescent microscopy images of cells 48 h post-transfection, expressing FUS-HA, NES-tdTomato-NLS (Std-Tomato) and either PDI or empty vector following immunocytochemistry in Neuro-2a cells. Cells transfected with 1 µm Mifepristone (row 1), FUS-HA-WT (row 2), mutant FUS-HA-R521G with empty vector (row 3) or mutant FUS-HA-R521G co-expressing PDI (row 4). Scale bar = 5 µm. (B) Quantification of the nuclear to cytoplasmic (N/C) fluorescence intensity ratio of Std-Tomato in Neuro-2a cells expressing FUS-HA-WT or mutant FUS-HA-R521G or co-expressing PDI in (A). Significantly more cytoplasmic reporter is present in cells expressing mutant FUS-HA-R521G (**p < 0.05) compared to FUS-HA-WT. However, co-expression of PDI (*p < 0.05) results in more reporter localised in the nucleus in FUS-HA-R521G cells compared to those expressing empty vector. Values represent mean ± SD, n = 3. **p < 0.01, *p < 0.05.