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. 2021 Aug 9;189(3):607–619. doi: 10.1007/s10549-021-06286-5

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — ARHGAP5-AS1 stabilizes SMAD7 via inhibition of SMAD7 ubiquitination. A The protein level of SMAD7 was detected by western blot in MDA-MB-231 cells and SKBR3 cells with knockdown of ARHGAP5-AS1. B The protein level of SMAD7 was detected by western blot in LM2 cells and BT549 cells with over-expression of ARHGAP5-AS1. C Knockdown of ARHGAP5-AS1 in MDA-MB-231 cells with treatment of 20 μM MG132. D Knockdown of ARHGAP5-AS1 in MDA-MB-231 cells with treatment of 40 μM CHX. E Immunoprecipitation was performed to enrich ubiquitin-SMAD7 complex in 293 T cells with indicated transfections. F Immunoprecipitation of FLAG was performed to enrich FLAG-tagged SMAD7 and HIS-tagged SMURF1/SMURF2 complex with or without over-expression of ARHGAP5-AS1 in 293 T cells. Each experiment was repeated three times independently