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. 2016 Mar 2;13:13. doi: 10.1186/s12977-016-0247-z

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© Zhang et al. 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 8 — Effect of 2-thio-6-azauridine on Vpu-induced ubiquitination of BST-2. 293T cells were co-transfected with plasmid pBST-2, pVphu and CW7, which express BST-2, Vpu and C-myc tagged ubiquitin respectively (1:2:4). 24 h post transfection, cells were treated with DMSO, 5 µM 2-thio-6-azauridine and/or 50 nM Concanamycin A for another 24 h. Cell lysates were then immunoprecipitated with rabbit BST-2 antibody and treated with PNGase F, which was used to remove N-glycans from BST-2, followed by immunoblotting with mouse C-myc and HA antibody for ubiquitin and BST-2 detection respectively. The normalized ubiquitination Level of BST-2 was shown as bar graph