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. 2022 Apr 28;13:2306. doi: 10.1038/s41467-022-30015-1

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© 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/.

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Fig. 7 — a Expression of frameshift variants in Drosophila eye tissue using GMR-GAL4 causes a rough eye phenotype. b Expression of frameshift variants in Drosophila muscle using MHC-GAL4 causes an abnormal wing posture phenotype. The percentage of flies with abnormal wing posture is plotted for 5 days after eclosion. n = 26, 26, 26, 32, and 35 flies for WT, D290V, N323Tfs*36, G328Afs*31, and G331Efs*28, respectively. c Adult flies expressing hnRNPA2 transgene using MHC-GAL4 were dissected to expose the dorsal longitudinal indirect flight muscle and stained with rhodamine-phalloidin (purple), hnRNPA2 (red), and DAPI (blue). hnRNPA2 WT localized exclusively to nuclei, whereas hnRNPA2 D290V accumulated extensively in cytoplasmic inclusions. Frameshift mutants showed both nuclear staining and diffuse sarcoplasmic accumulation. Scale bar, 10 μm. d hnRNPA2 expression in thoraces of adult flies driven by MHC-GAL4 in transgenic flies. Representative immunoblots from three independent experiments are shown. e Ratio of sarcoplasmic intensity of hnRNPA2 signal in indirect flight muscles. Error bars represent mean ± s.d. n = 12, 9, 12, 11, and 8 fly muscle samples for WT, D290V, N323Tfs*36, G328Afs*31, and G331Efs*28, respectively. ****P < 0.0001 by one-way ANOVA with Dunnett’s multiple comparisons test. f Thoraces of adult flies expressing hnRNPA2 transgene using MHC-GAL4 were dissected and sequential extractions were performed to examine the solubility profile of hnRNPA2. w1118 flies are included as a non-transgenic control. Representative immunoblots from three independent experiments are shown. g Quantification of RIPA-soluble and -insoluble fraction of hnRNPA2. Data represent mean ± s.d., n = 3 (WT, D290V, N323Tfs*36, G328Afs*31) and 2 (G331Efs*28) independent experiments. ***P = 0.0002 by two-way ANOVA with Dunnett’s multiple comparisons test.