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. 2021 Mar 2;5:14. doi: 10.1038/s41698-021-00153-8

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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. 5 — a Immunoprecipitates were analyzed with CYLD antibodies in HONE1 and HONE1-EBV cells. The arrow points to the destination band. Immunoprecipitates of RIP1 was taken as positive control. b 293T cells were transfected with Flag-CYLD and Myc-p18, and he cell lysates were subjected to immunoprecipitation with Flag antibodies, then the immunoprecipitates were probed with anti-Flag or anti-p18. Immunoprecipitates of Flag-CYLD and Myc-RIP1 was taken as positive control. c Proximity ligation assay indicating the interaction of CYLD and p18 in HONE1 cells (red: PLA positive signal; blue: DAPI, scale bar = 10 μm). d Endogenous CYLD (green) and p18 (red) in HONE1-NC and -shCYLD cells was visualized by immunofluorescence with anti-CYLD and anti-p18. DNA was stained with DAPI, and a merged view of the red and green channels within the same field is shown (merge) (scale bar, 10 μm). The interaction percent was calculated by Image J. e Schematic representation of the Flag-tagged full-length CYLD (FL) and its various deletion mutants, including N-terminal 1-303 deletion (1), Middle domain deletion (2), N-terminal (3) and C-terminal (4) constructs. f 293 T cells transfected with the indicated constructs were disrupted. The cell lysates were subjected to immunoprecipitation with anti-Flag. g Schematic representation of the Myc-tagged full-length p18 (FL) and its various deletion mutants, including N-terminal 1-34 deletion (1), N-terminal 36-66 deletion (2), C-terminal 68-99 deletion (3) and C-terminal 101-168 deletion (4) constructs. h 293 T cells transfected with the indicated constructs were disrupted. The cell lysates were subjected to immunoprecipitation with anti-Flag, and the immunoprecipitates were then probed with anti-Myc and anti-Flag. (Data are represented as mean ± SEM (n = 3). Differences were considered significant at ***p < 0.001).