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. 2018 Oct 23;9:4406. doi: 10.1038/s41467-018-06543-0

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© The Author(s) 2018

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. 1 — Components of LEC and SEC modulate TDP-43 toxicity. a TDP-43 expression causes eye degeneration that is suppressed by Ell RNAi and enhanced by Ell upregulation. Scale bars: external eye images (top), 100 μm; internal retinal section (bottom), 5 μm. Genotypes: control is gmr-GAL4(YH3)/+. gmr > TDP-43 is UAS-TDP-43/+; gmr-GAL4(YH3)/+. gmr > TDP-43 + Control RNAi is UAS-TDP-43/+; gmr-GAL4(YH3)/UAS-Control.RNAi^JF01355. gmr > TDP-43 + Ell RNAi is UAS-TDP-43/+; gmr-GAL4(YH3)/UAS-Ell.RNAi^HMS00277. gmr > TDP-43 + Ell is UAS-TDP-43/+; gmr-GAL4(YH3)/UAS-Ell^P{EP}G4098. b Quantification of retina thickness related to a. Three flies of each genotypes were measured (n = 3). Bars represent mean (SD). **P < 0.01, ****P < 0.0001 (two-tailed unpaired Student’s t-test). Genotypes are the same as indicated in a. A.U., arbitrary units. c Schematic of LEC and SEC. d Downregulation of SEC components (ear and lilli), or LEC component (Ice1) suppresses TDP-43 toxicity. Scale bars: external eye (top), 100 μm; internal retina section (bottom), 5 μm. Genotypes: gmr > TDP-43 is UAS-TDP-43/+; gmr-GAL4(YH3)/+. gmr > TDP-43 + ear RNAi is UAS-TDP-43/+; gmr-GAL4(YH3)/UAS-ear.RNAi^HMS00107. gmr > TDP-43 + lilli+/- is UAS-TDP-43/lilli^17–2; gmr-GAL4(YH3)/+. gmr > TDP-43 + Ice1 RNAi is UAS-TDP-43/UAS-Ice1.RNAi (SH09112.N from DRSC/TRiP); gmr-GAL4(YH3)/+. e Quantification of retina thickness related to d. Three flies of each genotypes were measured (n = 3). Bars represent mean (SD). **P < 0.01 (two-tailed unpaired Student’s t-test). Genotypes are the same as indicated in d. f TDP-43 expression in the adult neurons by elavGS causes climbing defects. Knockdown of Ell restores climbing ability. RU486 (8 mg/ml) was used to induce the expression of TDP-43 and Ell RNAi. EtOH was used as vehicle. Total of 100 flies were measured three times for each genotype at different time points. Bars represent mean (SD). ***P < 0.001, ****P < 0.0001 (two-way ANOVA followed by Tukey’s multiple comparison test). Significant differences are only indicated within the same time point. Genotypes: elavGS > TDP-43 is elavGS-GAL4, UAS-TDP-43/+. elavGS > TDP-43 + Ell RNAi is elavGS-GAL4, UAS-TDP-43/UAS-Ell.RNAi ^HMS00277. g TDP-43 protein levels in heads from 7, 14 and 21-day flies are not altered by Ell RNAi. n = 3 biological replicates. Tubulin was used as internal control. Bars represent mean (SD). NS, not significant (two-tailed unpaired Student's t-test with Welch’s correction). Genotypes and RU486 treatment are as in f