FIG 2.

Acetylation of ADA3 is counteracted by class III HDACs (SIRTs). (A) HEK293T cells were transfected with FLAG-ADA3 or empty vector. Thirty hours after transfection, cells were treated either with vehicle or 1 μM HDAC class I and II inhibitor trichostatin A (TSA) or 5 mM HDAC class III inhibitor nicotinamide (NAM) for 10 h. Cell lysates were then subjected to immunoprecipitation by M2 FLAG-agarose beads and immunoblotted with either anti-acetyl-lysine or FLAG-HRP antibodies. (B) The band intensities of acetylated FLAG-ADA3 over immunoprecipitated FLAG-ADA3 were quantified using ImageJ software and averaged from four independent experiments. The statistical significance between different groups was computed using Student's t test. NS, not significant; *, P ≤ 0.05.