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. 2021 Dec 13;18:38. doi: 10.1186/s12977-021-00584-y

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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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Trim28 decreases the expression of Tas protein and inhibits PFV replication through ubiquitin–proteasome pathway. a Trim28 down regulates the expression of Tas protein and the inhibition was reversed by MG132. HA-Tas combined with Flag-Trim28 or pCMV-Flag (as control) were transfected into HEK293T cells; 8 h before harvest, the cells were treated with DMSO or MG132 (5 μM). Western blot was performed to assess Tas protein levels; b Trim28 promotes Tas polyubiquitination. HA-tagged ubiquitin (HA-Ub), Myc-Tas combined with Flag-Trim28 were transfected into HEK293T cells, pCMV-Flag used as negative control. c MG132 rescues the suppression of PFV produced by overexpressed Trim28, and deletion of RBCC domain abrogates its inhibitory effect. HT1080 cells were transfected with Flag-Trim28, HA-Trim28-△RBCC or pCMV-Flag (as control) for 24 h and challenged with PFV (MOI = 0.1) for another 48 h. 8 h before harvest, one group of cells transfected with Flag-Trim28 were treated with MG132 (5 μM, another groups were treated with DMSO). d Deletion of RBCC domain of Trim28 cannot cause trigger the ubiquitination of Tas. In cellular ubiquitination assay, HEK293T cells were transfected with HA-tagged ubiquitin (HA-Ub), Myc-Tas combined with Flag-Trim28 or Flag-Trim28-△RBCC, pCMV-Flag used as negative control