FIG. 3.

Depsipeptide induces p53 acetylation. Representative Western blots show changes of p53 amount, in which A549 cells were treated with depsipeptide at different concentrations for 6 h (A) or at 0.1 μM for different incubation times after treatment (B). Proteins were extracted for Western blotting by using anti-p53 (P240 for panel A and DO-1 for panel B; these antibodies recognize full-length p53 and the N terminus of p53, respectively). A further RT-PCR assay was performed for testing changes in p53 mRNA at different intervals after treatment with depsipeptide (C). GAPDH is a loading control for the RT-PCR. (D) Cells were also treated with depsipeptide at different concentrations, and Western blotting was performed by using anti-p53 (Bp53-12, recognizing the unmodified C terminus). α-Tubulin, as a loading control, is shown under panels A, B, and D. (E) Cells were treated with depsipeptide at different concentrations, and protein was extracted for Co-IP by using antiacetyllysine antibody, followed by Western immunoblotting with anti-p53 (DO-1). ctr, control.