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. 2020 May 7;11(5):328. doi: 10.1038/s41419-020-2519-8

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© The Author(s) 2020

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. 1 — a, b Endogenous USP10 and HDAC6 interact with each other in H23 cells. a The anti-USP10 antibody or anti-IgG was used to immunoprecipitate USP10 in H23 cells followed by anti-HDAC6 Western blotting analysis (upper panel). The blot was stripped and reprobed with the anti-USP10 antibody (lower panel). b The reciprocal immunoprecipitation of a was performed. c USP10 physically binds to HDAC6. Bacterially-purified GST or GST-USP10 was incubated with His-HDAC6 isolated from E.coli, and the GST pull-down assays were performed followed by anti-HDAC6 Western blotting analysis (upper panel). His-HDAC6, GST, and GST-USP10 proteins were visualized via Coomassie blue staining (middle and lower panels). d, e Both the DAC1 domain and DAC2 domain of HDAC6 interact with USP10. d The schematic diagrams of HDAC6 full-length (FL) and deletion mutants. e The indicated vector and HDAC6 plasmids were co-transfected with HA-USP10 in 293T cells, followed by anti-Flag immunoprecipitation and subsequent anti-HA Western blotting analysis (upper panel). The blot was then stripped and reprobed with anti-Flag antibodies (middle panel). The input of HA-USP10 is shown in the lower panel. f, g The very C-terminus of USP10 binds to HDAC6. f The schematic diagrams of USP10 full-length (FL) and deletion mutants. g The indicated vector and USP10 plasmids were co-transfected with HA-HDAC6 in 293T cells, and anti-Flag immunoprecipitation was performed followed by anti-HA Western blotting analysis (upper panel). The blot was then reprobed with anti-Flag antibodies (middle panel). The input of HA-HDAC6 is shown in the lower panel.