FIG 3.

HDAC6 mediates the deacetylation of PA at Lys(664). (A) Schematic diagram of PA modification. NLS, nuclear localization signal. (B) 293T cells were transfected with Flag-PA or its acetylation dead mutants as indicated and then treated with tubacin (10 μM) for 17 h. Flag antibody was used to immunoprecipitate the wild type or acetylation dead mutants of Flag-PA, which were then analyzed by immunoblotting with the indicated antibodies. WCL, whole-cell lysates. (C) 293T cells were transfected with the indicated plasmids, followed by tubacin or DMSO treatment for 17 h. The cells were then treated with CHX (10 μg/ml) at the indicated time points. PA and actin were detected by immunoblotting with the indicated antibodies. (D) Calculated relative half-lives of PA, PA-K664R, and PA-K664Q, using the data from panel C. The percent intensity (log₁₀) was plotted versus time.