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. 2017 Jan 5;7:2129. doi: 10.3389/fmicb.2016.02129

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Copyright © 2017 Huang, Chen, Wang, Hsu, Tsai, Liu and Chang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Enhancement of Rta ubiquitination by TRIM5α. (A) 293T cells were cotransfected with plasmids expressing Rta, Flag-ubiquitin, and TRIM5α. Proteins in the lysates were immunoprecipitated (IP) using anti-Flag antibody and assessed by immunoblotting (IB) with anti-Rta antibody. (B) 293T cells were cotransfected with plasmids pFlag-Rta, pHA-Ub, and pLKO-shTRIM5α, which encoded Flag-Rta, HA-ubiquitin and shTRIM5α, respectively. Proteins in the lysates were immunoprecipitated (IP) using anti-Flag antibody and analyzed by immunoblotting (IB) with anti-HA antibody. At 24 h after transfection, cells were treated with 5 μM MG132 for additional 12 h to inhibit proteasome degradation. The asterisk indicates a non-specific band, also detected in previous studies (Chang et al., 2004; Yang et al., 2013). Ub-Rta, ubiquitinated Rta.