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. 2018 Dec 17;9:5342. doi: 10.1038/s41467-018-07718-5

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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. 4 — BAG3^P209L fails to collaborate with Hsp70 in client refolding. a Recombinant BAG3^P209L and BAG3^wt have normal affinity to HSPA8^NBD. b All of the BAG3 variants are capable of releasing fluorescent nucleotide from Hsc70/HSPA9. Results are the average of at least three experiments performed in triplicate. Error bars represent SD. See the methods for details. c, d Recombinant BAG3^P209L is not functional in promoting HSPA8 steady state ATP hydrolysis (c) and Hsp70-mediated refolding assays (d). Measurement of ATPase activity and denatured luciferase refolding was carried out in the presence of Hsc70, DnaJA2, and various concentrations of BAG3^WT or BAG3^P209L. Results are the average of at least three independent experiments performed in triplicate each. Error bars represent SD. e luciferase folding capacity of HEK293 cells expressing HSPB8 and BAG3^WT or indicated mutants of BAG3. Data represents the mean and standard deviation of two independent experiments (with three technical repeats for each experiment, Welch t test was used to calculate the P values, ** indicates P < 0.01). f Noncanonical interaction of BAG3^P209L with LVEAVY amyloid peptide. Source data are provided as a Source data file