Fig. 2. In vivo deacetylation of α-tubulin by HDAC6. (A) Effects of HDAC expression on tubulin acetylation in individual cells. NIH 3T3 cells grown on coverslips were transfected with HA-HDACs. After fixation, cells were immunostained with anti-acetylated α-tubulin and anti-HA antibodies. The cells transfected with HDAC6 are indicated by arrowheads. (B) Effects of HDAC expression on tubulin acetylation in TSA-treated cells. NIH 3T3 cells grown on coverslips were transfected with HA-HDACs and then treated with 100 nM TSA for 6 h. The cells transfected with HDAC6 are indicated by arrowheads. (C) Inhibition of the HDAC6-mediated tubulin deacetylation by a high concentration of TSA. NIH 3T3 cells grown on coverslips were transfected with HA-HDAC6 and then treated with 3 µM TSA for 6 h. The cells transfected with HDAC6 are indicated by arrowheads.