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. 2021 Jan 11;12:265. doi: 10.1038/s41467-020-20597-z

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© The Author(s) 2021

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. 3 — a Rescue of mtHSP70 solubility with protease-deficient LONP1. Triton X-100 extraction and Western blotting were used to analyze protein solubility in LONP1 knockdown cells re-expressing doxycycline (DOX)-inducible RNAi-resistant LONP1^WT and protease-deficient LONP1^S855A. b Protein aggregation in LONP1 knockdown cells re-expressing ATPase-deficient LONP1. Triton X-100 extraction and Western blotting were used to examine protein solubility in LONP1 knockdown 143B cells re-expressing doxycycline (DOX)-inducible RNAi-resistant LONP1^K529R. c Temporal features of mtHSP70 insolubility. mtHSP70 solubility was analyzed in 143B cells expressing DOX-inducible mtHSP70-HA. Cells were treated with DOX and 2 μM CDDO simultaneously or sequentially, as in Fig. 1f. d Comparison of OXA1L versus mtHSP70 solubility. 143B cells expressing DOX-inducible OXA1L-FLAG and mtHSP70-HA were treated with DOX and CDDO simultaneously or sequentially. Note the aggregation of mtHSP70 but not OXA1L in the sequential treatment. Scale bar, 10 μm. e Rescue of CDDO-induced mtHSP70 insolubility by CCCP or CHX. mtHSP70 solubility was analyzed in 143B cells expressing DOX-inducible mtHSP70-HA. Cells were treated with DOX and CDDO sequentially. CCCP or CHX was co-treated with CDDO as indicated.