Fig. 3.

GCN5L1 interacts with the canonical α-tubulin acetyltransferase αTAT1. (A,B) Co-immunoprecipitation studies revealed an association between GCN5L1 and αTAT1. 293T cells were co-transfected with GFP–αTAT1 plus control vector (Vector) or Flag–GCN5L1 (A), or co-transfected with Flag–GCN5L1 plus control vector (Vector) or GFP–αTAT1 (B), then anti-Flag (A), or anti-GFP (B) immunoprecipitates (IP) were subjected to immunoblot analysis with anti-Flag, anti-GFP and anti-α-tubulin antibody. α-Tubulin was used as loading control. (C) The interaction between GCN5L1 and αTAT1 was confirmed by the immunoprecipitation of endogenous GCN5L1 with Flag–αTAT1 versus an IgG control. (D,E) Confocal microscopy showed colocalization of GCN5L1 with αTAT1. HeLa cells were co-transfected with GFP–αTAT1 (green) and Flag–GCN5L1 (red) (D), or co-transfected with GFP–GCN5L1 (green) and Flag–αTAT1 (red) (E), then analyzed for the localization using anti-Flag antibodies. GFP fluorescence was visualized directly. DNA was visualized by DAPI staining (blue). Scale bars: 10 µm. The confocal images are a representative image of three independent experiments.