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. 2018 Mar 15;8:4630. doi: 10.1038/s41598-018-22879-5

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© The Author(s) 2018

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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TAF1 zinc knuckle and winged helix are imperative for effective cyclin promoter activity. (A) Incorporation of TAF1 proteins into TFIID. TFIID complexes were isolated by immunoprecipitation and incorporated HA-TAF1 variants detected by anti-HA immunoblotting. Additional TFIID subunits immunoprecipitated were detected by immunoblotting. The full-length blots are presented in Supplemental Figure S5. Quantitation of normalized HA-TAF1 and TAF protein levels are provided in Supplemental Table 2. (B) Chromatin immunoprecipitation of HA-TAF1 variants expressed in HEK293 cells followed by qPCR for cyclin D1 and cyclin A2 promoters (n = 4). (C) Luciferase assay of cyclin D1 and cyclin A2 promoter driven reporter constructs co-transfected with TAF1 variants. Luciferase activity was normalized for total protein and expressed relative to reporter activity without exogenous TAF1 (Empty), given a value of 1.0 (n = 3). All error bars represent standard deviation. Two-tailed analysis compared to WT with a 95% confidence, **p < 0.01, ***p < 0.0001 (Unpaired t-test).