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. 2021 Oct 15;12:6025. doi: 10.1038/s41467-021-26303-x

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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 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. 4 — a, f The G4C2 repeat RNA (primers detect C9orf72 intronic RNA) and act-1 mRNAs were assessed (RT-PCR) on mixed stage ΔC9 ^ubi (kasEx154), C9 ^ubi (kasEx262), eif-2A (gk358198); C9 ^ubi (kasEx262), and eif-2D (gk904876); C9 ^ubi (kasEx262) animals. b, g The nLuc and act-1 mRNAs were assessed by RT-PCR. c, h Worm lysates were processed for western blotting and immunostained with poly-GA and α-tubulin antibodies. d, i Quantification of poly-GA expression on western blots. The experiments were repeated four times in a, b, and d, and three times in f, g, and i, respectively, using 1 transgenic line per genotype (mean ± s.e.m.). One-way ANOVA with Dunnett’s multiple comparisons test was performed (e, j). e Lifespan of ΔC9 ^ubi (kasEx154), C9 ^ubi (kasEx262), and eif-2D (gk904876); C9 ^ubi (kasEx262) animals. Transgenic lines used: ΔC9^ubi (kasEx154), n = 75; C9^ubi (kasEx262), n = 72, eif-2D (gk904876); C9 ^ubi, n = 75). j Lifespan of ΔC9 ^neuro (kasEx159), C9 ^neuro (kasEx157), eif-2D (gk904876); C9 ^neuro (kasEx157), and eif-2D (gk904876) worms. Transgenic lines used: ΔC9 ^neuro (kasEx159), n = 62; C9 ^neuro (KasEx157), n = 63; eif-2D (gk904876); C9 ^neuro (kasEx157, n = 67; eif-2D (gk904876): n = 68). Mantel–Cox log-rank test was performed.