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. 2017 Sep 6;7:10657. doi: 10.1038/s41598-017-10728-w

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© The Author(s) 2017

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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Amt-87 dissociates P-TEFb from 7SK snRNP and promotes Tat-SEC formation at the HIV-1 promoter. (A) The HeLa-based F1C2 cells stably expressing the Flag-tagged CDK9 were treated with Amt-87 or DMSO for 24 hr. Nuclear extracts (NE) and anti-Flag immunoprecipitates (IP) derived from NE were analyzed by Western blotting for the indicated proteins and the results were quantified and shown on the right, with the numbers in lanes 1 and 3 set to 1. (B) NH1 cells transfected with the Tat-Flag-expressing plasmid were treated with Amt-87 (+) or DMSO (−) for 24 hr. NEs and anti-Flag IP derived from NE were analyzed by Western blotting for the indicated proteins. (C) Upon transfection with the plasmid expressing Tat-Flag for 24 hr, NH1 cells were treated with Amt-87 or DMSO for 24 hr and then subjected to ChIP-qPCR analysis to determine the occupancy of the indicated factors at three different locations (1–3) at and around the HIV-1 promoter region (diagramed on top). The ChIP signals were normalized to those of input and plotted, with the error bars indicating mean ± SD from three independent experiments.