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. 2018 Apr 16;8:6030. doi: 10.1038/s41598-018-24463-3

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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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3B12A scFv selectively degrades aggregated TDP-43 and the CMA signal further promotes degradation of aggregated TDP-43. (a,b) HaloTag pulse chase assay of TDP-43^WT and TDP-43^{mNLS,C173S/C175S}. (c,d) Quantified data for HaloTag pulse chase assay of (a,b) respectively. Each data point was obtained by normalisation to actin. 3B12A scFvs promoted the degradation of the TDP-43^{mNLS,C173S/C175S} mutant. Moreover, VH_VL-CMA significantly enhanced degradation compared with VH_VL. Differences were evaluated by two-way ANOVA (mean ± SD from three independent experiments; **p < 0.01, ***p < 0.005 and ****p < 0.001). N.S. indicates not significant. (e,f) Quantified data for HaloTag pulse chase assay of TDP-43^mNLS and TDP-43^C173S/C175S, respectively. Differences were evaluated by two-way ANOVA (mean ± SD from three independent experiments; *p < 0.05 and ***p < 0.005). N.S. indicates not significant. (g) HaloTag pulse chase assay for cytoplasmic aggregated TDP-43 in the presence of lactacystin or bafilomycin. (h) Quantitative analysis of (g) Each data point was obtained by normalisation to actin. Differences were evaluated by one-way ANOVA (mean ± SD from three independent experiments; **p < 0.01 and ***p < 0.005 vs DMSO control).