Fig. 1.

GCN5L1 deficiency impairs the acetylation of α-tubulin. (A,B) Reduced acetylated α-tubulin (Ac-Tub) levels are detected by immunoblot analysis in liver homogenates (A) and primary hepatocytes lysates (B) from GCN5L1-knockout (LKO) mice compared to WT mice. The antibody directed against α-tubulin (αTub) was used as loading control and the antibody directed against GCN5L1 to show the mouse genotype. Of note, a non-specific band (depicted with an arrowhead) was evident at a higher molecular mass than the specific GCN5L1 band. The accompanying histogram represents the relative (Rel) quantified ratio of ac-Tub to α-Tub normalized to the WT samples from three pairs of mouse livers. (C) GCN5L1-knockout causes reduced Ac-Tub levels in primary hepatocytes. Primary hepatocytes from WT and LKO mice were immunostained for endogenous Ac-Tub (green). DNA was visualized by DAPI staining (blue). Scale bar: 20 µm. (D,E) Overexpression of exogenous GCN5L1 restored Ac-Tub levels in LKO hepatocytes. WT and LKO hepatocytes were infected with control adenovirus (Ad-Ctrl) or with adenovirus coding for WT GCN5L1 (Ad-GCN5L1). (D) Ac-Tub levels were analyzed by immunoblotting. A quantification of the relative levels from three independent experiments is also shown. (E) Immunostaining as in C. Scale bar: 20 µm. Representative images are shown from three independent experiments. Data were expressed as mean±s.e.m. *P<0.05; **P<0.01 (unpaired Student's t-test).