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. 2022 Jul 1;144(3):413–435. doi: 10.1007/s00401-022-02450-3

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© The Author(s) 2022

Open AccessThis 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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Fig. 3 — NOVA1 expression is increased in ALS iPSC-MN and elevated cytoplasmic TDP-43 levels are associated with a reduction of NOVA1 mRNA levels. a–d Bar graphs of gene expression values for NOVA1 as reads per kilobase transcript per million mapped reads (RPKM) from a sALS and Ctrl generated in this study (n = 4 sALS, n = 4 Ctrl), b fALS and Ctrl generated in this study (n = 2 fALS, n = 3 Ctrl), c sALS and Ctrl from the AnswerALS consortium dataset (n = 9 sALS, n = 9 Ctrl), and d C9-ALS and Ctrl from the AnswerALS consortium dataset (n = 5 C9-ALS, n = 9 Ctrl). Data presented as mean ±SD. Statistical significance was determined by Mann-Whitney U test. a P value = 0.0286, b P value = 0.2000, c P value = 0.0315, d P value = 0.0599. e Western blot from total cell lysates of iPSC-MNs stained for NOVA1 and GAPDH. f Bar graph showing densitometric quantification of Western blot of NOVA1 normalized to GAPDH (n = 6 ALS; n = 6 Ctrl). Fold change ALS/Ctrl: 1.42. Data presented as mean ±SD. Statistical significance was determined by Mann-Whitney U test. P value = 0.0152. g Bar plot with NOVA1 expression values (in TPM) in iPSC-MN with control siRNA or siRNA targeting TARDBP (GEO ID: GSE121569; n = 6 control siRNA, n = 6 TARDBP siRNA). Data presented as median +IQR. Statistical significance was determined by Mann-Whitney U test. P value = 0.3095. h Bar plot with NOVA1 expression values (in TPM) in SH-SY5Y with control siRNA or siRNA targeting TARDBP (GEO ID: GSE122069; n = 3 control siRNA, n = 3 TARDBP siRNA). Data presented as median +IQR. Statistical significance was determined by Mann-Whitney U test. P value = 0.4000. i Bar plot with NOVA1 expression values (in RPKM) in mouse striatum with control ASO or ASO targeting TARDBP (GEO ID: GSE27394; n = 4 control ASO, n = 5 TARDBP ASO). Data presented as median +IQR. Statistical significance was determined by Mann-Whitney U test. P value = 0.9048. j Bar plot with NOVA1 expression values (in RPM) in mouse brain without and with TDP-43-dNLS expression (GEO ID: GSE65973; n = 4 Ctrl, n = 4 TDP-43-dNLS). Data presented as median +IQR. Statistical significance was determined by Mann-Whitney U test. P value = 0.0286. k Bar plot with NOVA1 expression values (in DESeq2 normalized counts) in iPSC-MN (4 Ctrl, 4 HNRNPA2B1 mutant lines) without and with puromycin stress (GEO ID: GSE157467; n = 8 untreated, n = 8 puromycin). Data presented as median +IQR. Statistical significance was determined by Mann-Whitney U test. P value = 0.0379