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. 2021 Sep 16;12:5488. doi: 10.1038/s41467-021-25809-8

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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. 8 — a Upper panel: IB of wild-type (ctrl), Hecw^KO (KO) and RFP-Hecw-overexpressing (elav>RFP Hecw) fly heads with the indicated antibodies. Profilin levels increase in Hecw^KO and decrease upon Hecw overexpression. Values reported below each blot are normalised for tubulin and represent the fold change relative to the control, presented as mean of three experiments. Bottom panel: mRNA level of profilin was measured by qPCR in the adult heads of the indicated genotypes. The reported expression levels relative to control are expressed as mean ± SD calculated over three experiments. Ns, not significant by unpaired two-tailed t-test. b IF analysis of adult brains from 1-day-old flies with the indicated antibodies (n = 2). Neurons are marked with the anti-elav antibody (purple); red, Hecw; blue, DAPI. Scale bar, 5 μm. c IF analysis of adult brains from 1-day-old flies of the indicated genotypes, not treated (nt) or treated with 1,6 hexanediol (1,6 hex). (n = 3). Scale bar, 5 μm. d Quantification of RNP size in fly neurons of the indicated genotypes and conditions. RNP size is significantly increased in Hecw^KO flies (KO;Me31B::GFP **P < 0.01) and Hecw^CI (CI;Me31B::GFP **P < 0.01). 1,6 hexanediol treatment significantly reduces RNP size in control flies. Size mean ± SD is reported. ****P < 0.0001 by two-tailed Mann–Whitney test (ten heads/genotype from two biological replicates). e Profilin and Orb levels are restored upon concomitant Hecw overexpression and Fmr1 silencing. Left panel: Profilin expression in fly brains of the indicated genotypes. Values reported below each blot are normalised for tubulin and represent the fold change relative to the control, presented as mean of three experiments. Right panel: Orb expression in fly ovaries of the indicated genotypes. Values reported below each blot are normalised for tubulin and represent the fold change relative to the control, presented as mean of two experiments.