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. 2023 Apr 7;8:142. doi: 10.1038/s41392-023-01354-2

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© The Author(s) 2023

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 — OTUB1 cooperates with SMURF1 to regulate the ubiquitination and stability of FGFR2. a Immunoprecipitates of Nedd4 family proteins in HEK293T cells transfected with Myc-Fgfr2 and immunoblotted with the indicated antibodies. b Examination of in vivo interaction between SMURF1 and OTUB1 in HEK293T and osteoblast cells. c Immunoprecipitates of FGFR2 ubiquitination in HEK293T cells transfected with Flag-Smurf1-WT, Flag-Smurf1-CA, and Flag-Cbl, and immunoblotted with the indicated antibodies. d Immunoprecipitates of FGFR2 ubiquitination in HEK293T cells transfected with Flag-Smurf1 or indicated Flag-Otub1 (WT, D88A, C91S, H265A, and A/S/A) and immunoblotted with the indicated antibodies. e Immunoblot analysis of the interaction between SMURF1 and its substrate FGFR2 in HEK293T cells with or without GFP-Otub1. f Immunoblot analysis of the interaction between SMURF1 and its E2 UbcH5c in vitro. g Immunoblot analysis of the ubiquitination of FGFR2 in bones from Smurf1^+/+ and Smurf1^−/− mice. h Immunoblot analysis of FGFR2 expression in Smurf1^+/+ and Smurf1^−/− osteoblast (OB) cells. i Immunoblot analysis of FGFR2 expression in Smurf1^+/+ and Smurf1^−/− bones. j Immunoblot of the signaling proteins including PI3K and MAPK signaling in Smurf1 WT and KO osteoblast cells with 20 ng/ml FGF10 stimulation