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. 2018 Feb 10;55(9):7533–7552. doi: 10.1007/s12035-018-0881-7

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© The Author(s) 2018

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits use, duplication, 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.

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Fig. 3 — ced-10 decreases α-SYN inclusions in C elegans. a Representative confocal pictures obtained from animals containing the genomic array pkIs2386 [Punc-54_::α-SYN::YFP] expressing α-SYN in body wall muscle cells at L4 + 5 days of development (7 days post hatching). Green staining in all figures represents α-SYN::YFP inclusions in muscle cells. The vulva (V, thick arrow) was used as a reference to analyze the same central section in all worms. A representative area was highlighted and expanded in each panel, to better visualize the α-SYN::YFP accumulation. a (first panel) α-SYN inclusions were detected in a C. elegans model of α-SYN miss folding in which α-SYN is expressed under the control of the unc-54 promoter. a (second panel) α-SYN apparent aggregates are increased in ced-10(n3246) mutant nematodes. a (third panel) CFP::CED-10 expression (array baEx167 [Pced-10::CFP::CED-10]) decreased the number of α-SYN inclusions in a ced-10(n3246) background. a (fourth panel) The blue fluorescence marker (CFP) represented the endogenous expression of CED-10 in a ced-10(n3246) background for rescuing α-SYN accumulation. Magnification bar is 10 μm. b Quantification of the number of α-SYN inclusions per area. Data are mean ± SEM. Between 30 and 35 animals were analyzed per genotype. Three different transgenic lines expressing CFP::CED-10 were generated and analyzed independently. Statistics: one-way ANOVA with a Tukey post hoc test. ***P < 0.005. c The movement of YFP::α-SYN animals is hampered by the mutation ced-10n(3246). Thrashing behavior (bends/min) was video recorded and the resulting images were analyzed by the ImageJ software. Data are mean ± SEM. Between 20 and 30 animals were recorded per experiment and the same experiment was repeated 3 independent times. d Immunoblotting analysis of protein extracts from 5 days post L4 old YFP::α-SYN synchronized animals, using anti-α- SYN antibody without and with the ced-10(n3246) mutation (wild type and ced-10 respectively). The amount of α-SYN insoluble species was increased by the ced-10(n3246) mutation.