Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Aug 9;12(4):261–278. doi: 10.1007/s13238-020-00751-5

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

HDAC3 directly enhances the TBK1 kinase activity via its deacetylase activity. (A and B) Luciferase IFN-β reporter activity analyzing (A) and Immunoblot analysis of phosphorylated and total TBK1 or IRF3 (B) in HEK293T cells plus RGFP966 in dose (50, 100, 200 nmol/L), infected with SeV for 10 h. (C) Immunoblot analysis of HEK293T cells (1.5 × 10⁶) transiently co-transfected for 48 h with HA-tagged TBK1 together with vector encoding Flag-tagged wild-type HDAC3, HDAC3 (1–316 aa), or HDAC3 (317–428 aa) (above lanes), assessed before (10% input) or after (IP) immunoprecipitation with anti-Flag antibody. (D) Immunoblot analysis of HEK293T cells (1.5 × 10⁶) transiently co-transfected for 48 h with HA-tagged HDAC3 with vector encoding EGFP-tagged wild-type or mutant TBK1 (above lanes), assessed before (10% input) or after (Co-IP) Co-immunoprecipitation with the indicated antibodies. (E) TBK1 kinase activity assay combined with purified GST-HDAC3. HEK293T cells (4.0 × 10⁵) was transfected with vector or Flag-tagged TBK1. The expressed proteins immunoprecipitated by Flag tag antibodies and purified GST-HDAC3 or BSA were introduced into a mixture as indicated for TBK1 kinase activity assay. (F) In vitro kinase assay. HEK293T cells (4.0 × 10⁵) were transfected with Flag-tagged TBK1, HA-tagged IRF3 and Flag-tagged HDAC3 separately (left). The expressed proteins were immunoprecipitated by the indicated tag antibodies and incubated together as indicated. The GST-TBK1 purified protein and the peptide of IRF3 full length protein with kinase activity were introduced into a mixture containing recombinant purified HDAC3 (right). Data are representative of three independent experiments. Graphs show mean ± SD; n = 3. *P < 0.05; (Student’s t-test)