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. 2019 Jun 11;9:8462. doi: 10.1038/s41598-019-44934-5

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© The Author(s) 2019

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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TARDBP assembles protein complexes that support the lifecycle of HBV. (a) An experimental design for the identification of TARDBP-interacting proteins. In the first step, nuclear lysates were prepared from the HBV producing cell line T23, immunoprecipitated by a TARDBP antibody and subjected to LC-MS/MS analysis. A literature search was done on the top scorers to ascertain which of them likely played a role in the HBV life cycle. In the second step, interactions of the candidate proteins in vivo were validated by western blotting using the same cell line as in (a) but with an exogenously expressed TARDBP protein. (b) The set of 8 proteins that scored a coverage of >10 on the LC/MS-MS analysis and were found from literature to have a role in HBV replication. The figure includes the specified role and the literature associated with each protein. (c) Nuclear lysates of the T23 cells expressing FLAG-tagged TARDBP were precipitated by the anti-FLAG antibody or the control mouse IgG and subjected to western blotting for each protein using their specific antibodies as shown.