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. 2023 Jul 24;14:4450. doi: 10.1038/s41467-023-39700-1

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© The Author(s) 2023

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. 2 — a 14-3-3(FTT-2) associates with BAG3-bound Hsc70/(HSP-1); Hsc70(HSP-1) recognizes and binds ubiquitinylated aggregates, which are transported via microtubules to the aggresome-like compartment. C. elegans gene names are indicated. b-c, j-l Solid white lines outline the soma cell body; white dashed lines outline the nucleus; scale bar = 2 µm. b Touch neuron expressing bzIs166[P_mec-4mCherry]; bzSi3[P_mec-7GFP::IFD-1]. Left: Control Ad2 DMSO; Right: Ad2 ALM, 5 mM colchicine in DMSO added from L4 to Ad2. Images are representative of N > 90 ALMs. c AID dynein heavy chain knockdown strain expresses bzIs166[P_mec-4mCherry]; bzSi6[P_mec-7TIR1::TagBFP]; bzSi3[P_mec-7GFP::IFD-1]; dhc-1(ie28[DHC-1::degron::GFP]); control strain lacks TIR1; 5 mM auxin exposure from L4 to Ad2. Images are representative of N > 90 ALMs. d-i. Data are 3 trials, two-tailed t test, error bars SEM. d Number of IFD-1-positive puncta per Ad2 ALM in bzIs166[P_mec-4mCherry]; bzSi3[P_mec-7GFP::IFD-1]. Control DMSO; colchicine 5 mM, L4 to Ad2 exposure. N = 37 and 27 ALM, respectively; ****P = 0.0001. e Diameter (µm) of IFD-1-positive foci in Ad2 ALM. Strain and treatment are the same as in (b). Ad2 N = 101 and 143 ALM respectively; ****P = 0.0001. f Percentage of juxtanuclear foci. Strain and treatment are the same as in (b). N = 37 and 27 ALM respectively, ****P = 0.0001. g Number of IFD-1-positive puncta per Ad2 ALM. Strains are bzIs166[P_mec-4mCherry]; bzSi3[P_mec-7GFP::IFD-1]; dhc-1(ie28[DHC-1::degron::GFP]) and bzIs166[P_mec-4mCherry]; bzSi6[P_mec-7TIR1::TagBFP]; bzSi3[P_mec-7GFP::IFD-1]; dhc-1(ie28[DHC-1::degron::GFP]); auxin exposure from L4 to Ad2. N = 35 and 25 ALM respectively, *P = 0.019. h Diameter (µm) of IFD-1-positive foci in Ad2 ALM. Strains are the same as in (c), with control –/– lacking P_mec-7TIR1 and both strains exposed to 5 mM auxin from L4 to Ad2. N = 98 ALM and 48 ALM respectively, **P = 0.0011. i Percentage of juxtanuclear foci for strains presented in (c). N = 36 ALM and 31 ALM respectively, **P = 0.0041. j-l Boxes indicate the range of coefficient. j Representative of adult touch neuron expressing bpIs151[P_sqst-1SQST-1::GFP+unc-76(+)]; bzEx261[P_mec-7RFP::IFD-2]. Quantification of colocalization of N = 10 ALMs, graphed on the right as Mander’s overlap coefficient of mScarlet channel over GFP channel. Range minimum (0.46), maximum (0.99), and mean (0.82) coefficient. k Representative of adult touch neuron expressing bzSi51[P_mec-7FTT-2::mSc]; bzIs3[P_mec-7GFP::IFD-1]. Quantification of colocalization of N = 10 ALMs, graphed on the right as Mander’s overlap coefficient of GFP channel over mScarlet channel. Range minimum (0.17), maximum (0.95), and mean (0.73) coefficient. l Representative of adult touch neuron expressing bzSi53[P_mec-7mSc::HSP-1]; bzIs3[P_mec-7GFP::IFD-1]. Quantification of colocalization of N = 10 ALMs, graphed as Mander’s overlap coefficient of GFP channel over mScarlet channel. Range minimum (0.24), maximum (0.99), and mean (0.77) coefficient.