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. 2019 Mar 20;10:1288. doi: 10.1038/s41467-019-09270-2

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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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Fig. 3 — XPB recruits KAT2A-containing HAT complexes to chromatin. a TFIIH-XPB^WT was immunoprecipitated from nuclear extracts of stably transfected cells using anti-GFP antibodies (GFP-Trap) and washed with 200 mM salt (Lanes 2 and 4). Control IPs were performed with anti-GFP on an XP-B/CS^F99S cell line stably expressing GFP alone (Lanes 1 and 3). Proteins on the resin were resolved by SDS-PAGE and immunoblotted using polyclonal rabbit anti-GFP, monoclonal mouse anti-p62, polyclonal rabbit anti-CDK7, polyclonal rabbit anti-KAT2A (SCBT) (Supplementary Figure 9), polyclonal rabbit anti-KAT2B, polyclonal rabbit anti-WDR5, or polyclonal rabbit anti-SUPT7L antibodies. Source data are provided as a Source Data file. b The U2OS17 cell line was transiently transfected with 1 μg of expression vectors for the following proteins: LacR-GFP, XPB^WT-LacR-GFP, XPB^F99S-LacR-GFP, or XPB^320–782-LacR-GFP together with 1 μg of expression vector for Flag-KAT2A. Colocalization of KAT2A with GFP was detected by immunofluorescence staining using a polyclonal rabbit anti-flag antibody. The values on the graph represent the percentage of colocalization of KAT2A with GFP on the array (error bars represent SD from three independent quantifications). c Purified wild-type recombinant core TFIIH (cIIH-XPB^WT) was incubated with purified Flag-KAT2A (400 ng) and pull-down assays were performed using either unspecific anti-IgG or polyclonal rabbit anti-KAT2A (SCBT) antibody (Supplementary Figure 9). After washing, proteins on the resin were resolved by SDS-PAGE and immunoblotted using mouse monoclonal anti-XPB or anti-p52 antibodies (two subunits of TFIIH) or monoclonal mouse anti-KAT2A (IGBMC) antibody. Source data are provided as a Source Data file. d Bacterially expressed recombinant KAT2A (rKAT2A) (400 ng) was tested for its ability to interact with either rXPB^WT (lanes 1–3) or rXPB^F99S (lanes 4–6) (500 ng) in a pulled-down assay (IP-XPB). After washing, proteins on the resin were resolved by SDS-PAGE and immunoblotted using mouse monoclonal anti-XPB and mouse monoclonal anti-KAT2A antibodies (IGBMC). Lanes 1 and 4, immunoprecipitations with an irrelevant antibody (IP-IgG). Source data are provided as a Source Data file