Respective secondary antibodies

Western blotting was carried out as described previously (7 (link)). Briefly, SMMC-7721 hepatoma cells were treated with various concentrations (5, 25, 50, or 100 µM) of PTER for 24 h. PTER (Sigma, St. Louis, MO, USA) was dissolved in dimethyl sulphoxide (DMSO); DMSO alone was used to treat cells in the control group. After washing with PBS, cells were then lysed using RIPA buffer, which was mixed with the Halt Phosphatase and Protease Inhibitor Cocktail (Thermo Fisher Scientific, Inc., Waltham, MA, USA). After the protein concentration was quantified, 70 µg of protein was resolved by SDS-polyacrylamide gel electrophoresis (PAGE) and transferred to a PVDF membrane using an Electrophoresis Transfer System (Bio-Rad). Subsequently, membranes placed on a swing table were blocked with TBS-Tween and 5% dry milk and were then probed with anti-MTA1, p53 or Ac-p53 (Abcam, Cambridge, UK) primary antibodies overnight at 4°C. Respective secondary antibodies (Abcam) were used at 1:1,000–1:2,000 (v/v) dilutions in TBS-Tween and 5% fat-free powdered milk for 1 h at room temperature; β-actin antibodies were used as a loading control. Finally, blots were visualized by SuperSignal West Pico enhanced chemiluminescence substrate (Thermo Fisher Scientific, Inc., Waltham, MA, USA), and the density of the blots was quantified using Image J Software (Bio-Rad).