Figure 7.

Lys692 deacetylation controls the dimerization of TBK1. (A) Immunoblot analysis of endogenous acetylation of TBK1 at Lys692 (Ac-K692) in HEK293T cells (1.5 × 10⁷), followed by infection with SeV for 0–9 h, before co-immunoprecipitation (with anti-TBK1 or IgG as a control) and immunoblot analysis with anti-Ac-K692, anti-phosphorylated and total TBK1or IRF3. (B) In vitro deacetylation assay. Assays were performed as in Fig. 6B. Immunoblot analysis with antibody to TBK1 acetylated at Lys692 (Ac-K692), anti-Flag, anti-HDAC3. (C) Effect of HDAC3 overexpression on TRAF3-TBK1, AZI2-TBK1 association. Immunoblot analysis of HEK293T cells (1.5 × 10⁶) transiently co-transfected for 48 h with the indicated plasmids before co-immunoprecipitation (with anti-Flag or IgG as a control) and immunoblot analysis were performed with the indicated antibodies. (D) Immunoblot analysis of TBK1 in dimerization in HDAC3^fl/fl or HDAC3^fl/fl Lyz2-Cre BMDMs and left infected with VSV for the indicated times, followed by native PAGE. (E) Immunoblot analysis of TBK1 in dimer or monomer form in HEK293T TBK1-knockout cells (4 × 10⁵), transiently transfected for 48 h with Flag-tagged empty vector (VEC), wild type (WT) or TBK1 mutants (K692Q or K692R), followed by native PAGE. Data are representative of three independent experiments