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. 2021 Jun 23;4:778. doi: 10.1038/s42003-021-02303-x

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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. 6 — a The effect of AKT inhibitor (MK2206) or 26S proteasomal inhibitor MG132 on TSPYL5 expression in H460 cells transfected with TSPYL5 (WT, mutant T120A or T120D). VC: pcDNA3.1-transfected control. The activity of AKT inhibitor was confirmed by Western blot analysis of p-AKT. b In vivo ubiquitination assay of H460 cells transfected with HA ubiquitin and TSPYL5 or related mutants (T120A, T120D, and T409A) and treated with or without MK2206. Cells were lysed and immunoprecipitated with anti-TSPYL5 antibody. Ubiquitinated TSPYL5 was determined with anti-HA antibody. c Treatment of A549 cells with the SUMO1 modification inhibitor ginkgolic acid inhibited nuclear translocation of TSPYL5. A549 cells attached on coverslips were treated with ginkgolic acid (3 μM, 4 h), and intracellular staining of TSPYL5 was performed after fixation and permeabilization. Scale bar: 20 μm. d In vivo SUMOylation assay of H460 cells transfected with His₆-SUMO1 and TSPYL5 or related mutants T120A. Cell lysates were precipitated with Ni-agarose bead and Western blot analysis was performed with anti-TSPYL5 antibody.